CN113462812B - 一种苹果优质抗病基因组辅助预测方法及其应用 - Google Patents
一种苹果优质抗病基因组辅助预测方法及其应用 Download PDFInfo
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Abstract
本发明提供了一种苹果优质抗病基因组辅助预测方法及其应用，属于植物鉴别与育种技术领域，所述苹果优质抗病基因组辅助预测方法依赖于分子标记实现，所述分子标记共计319个，包括的318个SNP标记和1个InDel标记；所述分子标记与苹果属植物果实成熟期、果形、果面着色度、平均单果重、可溶性固形物含量、果汁pH值、苹果酸含量、绿原酸含量、原花青素B2含量、采收时果肉硬度、采收时果肉脆度、果肉硬度保持、果肉脆度保持、果实轮纹病抗病性、炭疽叶枯病抗病性和短枝型等16个性状相关联。本发明的分子标记能够用于苹果种质资源评价和育种，能够极大地提高苹果育种效率，缩短育种周期。
Description
技术领域
本发明涉及植物鉴别与育种技术领域技术领域，尤其涉及一种苹果优质抗病基因组辅助预测方法及其应用。
背景技术
苹果是我国重要的经济作物之一，也是世界重要的栽培树种之一。目前我国苹果总产量占世界的50％以上。但是我国苹果品种单一，‘红富士’占我国苹果总产量的70％以上。因此，选育多样性品种，改善我国苹果品种结构，满足苹果多元化市场需求，调整产业结构是保障苹果产业健康发展的迫切需要。
长期以来，苹果育种的主要通过杂交育种和芽变选种等途径来改良旧品种或选育新品种，但由于苹果实生树童期长，基因型高度杂合，自交不亲和，以及缺少可靠的早期选育择方法，使得苹果育种效率难以提高。随着现代生物育种技术的发展，以标记辅助选择(Marker assisted selection,MAS)和全基因组选择(Genomic selection，GS)为代表的分子育种技术相继建立并逐渐应用于生物育种实践，为实现性状的早期选择，缩短世代间隔，压缩群体规模，提高育种精确度提供了可行的技术解决方案。但是，MAS采用单个或少数标记直接选择有益基因型或基因型组合，适用于单基因或主基因控制的质量性状，但苹果质量性状不多，MAS应用范围不大。GS采用覆盖全基因组的高密度SNP芯片或全基因组测序(Next generation sequencing，NGS)进行标记分型，通过估算效应值，选择预测育种值(Genomic estimated breeding value，GEBV)。GS适用于多基因数量性状，但高密度芯片和NGS成本偏高，苹果育种群体往往较大，GS应用很受限制，加上苹果的重要经济性状非加性遗传比例高，GS对于非加性遗传的预测准确性较低。
发明内容
本发明的目的在于提供一种苹果优质抗病基因组辅助预测方法及其应用。
为了实现上述发明目的，本发明提供以下技术方案：
本发明提供了苹果属植物性状相关分子标记，包括表1中的分子标记中的一种或几种；
表1苹果属植物性状相关分子标记
本发明还提供了上述方案所述分子标记的每个基因型对相应性状的基因型效应值或基因型组合效应值，如表2所示：
表2每个分子标记的每个基因型对相应性状的基因型效应值
本发明还提供了检测上述方案所述分子标记的引物组合，包括SEQ ID NO.1～SEQID NO.638所示的引物组合。
本发明提供了一种上述方案所述分子标记的检测方法，包括以下步骤：
1)提取待测苹果属植物的基因组DNA；
2)采用上述方案所述引物组对待测苹果属植物的基因组DNA进行多重PCR扩增，得到扩增产物；
3)采用二代测序方法测得所述扩增产物的基因型，得到待测苹果属植物样本的基因型。
优选的，步骤2)中所述多重PCR扩增的反应体系以30μL计，包括以下组分：上述方案中所述引物组合8μL、MP004_Cu Panel Mix 8μL、DNA 50～200ng、3xT酶10μL和余量的H2O；所述引物组合中每条引物的浓度为0.24μM；所述多重PCR扩增的反应程序包括：95℃3min；95℃30s，60℃4min，16个循环；72℃4min延伸所述多重PCR扩增的反应程序包括：95℃3min；95℃30s，60℃4min，16个循环；72℃4min延伸。
优选的，步骤3)中所述二代测序深度为1200x。
本发明提供了一种判定苹果属植物样本的性状表型或者计算苹果属植物样本的性状的预测表型值的方法，包括以下步骤：
上述方案所述的分子标记对应的性状的群体平均值，分别为：果实成熟期159.45DAFB，果面着色度56.35％，平均单果重106.63g，可溶性固形物含量14.85％，果汁pH3.34，苹果酸含量5.83mg/mL，采收时果肉硬度12.18kg/cm2，采收时果肉脆度1.31kg/cm2，果肉硬度保持2.41月、果肉脆度保持2.19月，果实轮纹病抗病性21.34mm，短枝型0.99；
按照上述方案所述的方法得到待测苹果属植物样本的基因型；根据所述待测苹果属植物样本的基因型，按照所述的群体平均值和上述方案所述的基因型效应值或基因型组合效应值，利用如下标准判定待测苹果属植物样本的性状表型或者使用如下预测模型计算待测苹果属植物样本的性状的预测表型值：
所述标准包括：
(1)炭疽叶枯病抗病性：
当S1202的基因型为CC且zhwy64的基因型为CC时，判定为抗病；其它基因型判定为感病；
(2)果形：
当newdy202的基因型为CC、SIZE2270的基因型为GG、SIZE5253的基因型为CC、SIZE9100的基因型为GG或者SIZE9195的基因型为AA时，判定果形为圆锥-圆；
当SP031的基因型为CC、SP081的基因型不为CT或者XDY231的基因型为GG时，判定果形为扁圆-圆；
所述预测模型包括：
(3)绿原酸含量或原花青素B2含量采用基因型组合模型；
按照绿原酸的分子标记的基因型或原花青素B2含量的分子标记的基因型组合估算效应值，用基因型组合效应值建立GAP模型，公式为：
GPV＝α×(GcE+μ)+β；
GPV为预测的表型值；GcE为该性状各个标记的基因型组合效应值；μ为训练群体该性状的表型均值；α和β分别为线性回归系数和剩余参数；
(4)果实成熟期、可溶性固形物含量、果汁pH值、采收时果肉硬度、采收时果肉脆度、果肉硬度保持性、果肉脆度保持性或果实轮纹病抗病性采用加性模型，公式为：
k
GPV＝α×(∑GE+μ)+β
i＝1
GPV为预测的表型值；GE为该性状标记的基因型效应值；k为该性状的标记数；μ为训练群体该性状的表型均值；α和β分别为线性回归系数和剩余参数；
平均单果重、苹果酸含量、果面着色度和短枝型采用固定效应模型，预测公式为：
k
GPV＝α×(Fx+γ×∑GnE+μ)+β
i＝1
GPV为预测的表型值；Fx为固定效应标记基因型效应值；GnE为该性状非固定效应标记的基因型效应值；k为该性状的非固定效应的标记数；μ为训练群体该性状的表型均值；γ为缩值系数；α和β分别为线性回归系数和剩余参数；
平均单果重的固定效应为：XDY160的基因型为AA或SIZE4849的基因型为GG或SIZE4161的基因型为GG，Fx分别为-104.8，-100.7和-101.4；
苹果酸含量的固定效应为Ma、MA202和SAUR-5的基因型组合效应值；
果面着色度的固定效应分别为ZZZ162与zwy6和ZZZ162与color1245的基因型组合效应值；
短枝型的固定效应为neww45与S1245和neww45与ww19的基因型组合效应值；
本发明还提供了上述方案所述苹果属植物性状相关分子标记或者所述基因型效应值或基因型组合效应值或者所述引物组的如下一种或几种应用：
1)苹果属植物性状鉴定；2)苹果属植物指纹图谱或分子身份证构建；3)苹果属种质资源基因型鉴定；4)苹果属植物杂交育种；5)苹果属植物新品种分子DUS测试；
所述苹果属植物杂交育种包括：杂交亲本的选择与杂交组合的选配、杂交世代数设计和杂种后代分子辅助选择中的一种或几种。
本发明提供了苹果属植物性状相关分子标记，所述分子标记共计319个，包括的318个SNP标记和1个InDel标记；所述分子标记与苹果属植物果实成熟期、果形、果面着色度、平均单果重、可溶性固形物含量、果汁pH值、苹果酸含量、绿原酸含量、原花青素B2含量、采收时果肉硬度、采收时果肉脆度、果肉硬度保持、果肉脆度保持、果实轮纹病抗病性、炭疽叶枯病抗病性和短枝型等16个性状相关联。本发明的分子标记能够用于苹果种质资源评价和育种，能够极大地提高苹果育种效率，缩短育种周期。
具体实施方式
本发明提供了苹果属植物性状相关分子标记，包括如前文表1所示的分子标记中的一种或几种。
在本发明中，所述分子标记包括318个SNP标记和1个InDel标记。
在本发明中，所述SNP分子标记基于黄瓜基因组序列信息版本号是GDDH13 v1.1。
本发明还提供了上述方案所述分子标记的每个基因型对相应性状的基因型效应值或基因型组合效应值，如前文表2所示。
本发明还提供了检测上述方案所述分子标记的引物组，包括SEQ ID NO.1～SEQID NO.638所示的引物组合。在本发明中，所述引物组对应检测的标记的名称、上游引物和下游引物如表3所示：
表3检测不同分子标记的引物组
本发明还提供了一种检测上述方案所述分子标记的方法，包括以下步骤：
1)提取待测苹果属植物的基因组DNA；
2)采用上述方案所述引物组对待测苹果属植物的基因组DNA进行多重PCR扩增，得到扩增产物；
3)采用二代测序方法测得所述扩增产物的基因型，得到待测苹果属植物样本的基因型。
本发明首先提取待测苹果属植物的基因组DNA。
本发明对提取待测苹果属植物的基因组DNA的方法没有特殊限制，采用本领域的常规方法即可。
得到待测苹果属植物的基因组DNA后，本发明采用上述方案所述引物组对苹果属植物待测样本基因组DNA进行多重PCR扩增，得到扩增产物。
在本发明中，所述多重PCR扩增的反应体系以30μL计，优选的包括以下组分：上述方案所述引物组合8μL、MP004_Cu Panel Mix 8μL、DNA 50～200ng、3xT酶10μL和余量的H2O；所述引物组合中每条引物浓度优选为0.24μM；所述多重PCR扩增的反应程序优选的包括：95℃3min；95℃30s，60℃4min，16个循环；72℃4min延伸。
得到扩增产物后，本发明采用二代测序方法测得所述扩增产物的基因型，得到待测苹果属植物样本的基因型；所述二代测序的深度优选为1200x。
本发明还提供了一种判定苹果属植物样本的性状表型或者计算苹果属植物样本的性状的预测表型值的方法，包括以下步骤：
上述方案所述的分子标记对应的性状的群体平均值分别为：果实成熟期159.45DAFB，果面着色度56.35％，平均单果重106.63g，可溶性固形物含量14.85％，果汁pH3.34，苹果酸含量5.83mg/mL，采收时果肉硬度12.18kg/cm2，采收时果肉脆度1.31kg/cm2，果肉硬度保持2.41月、果肉脆度保持2.19月，果实轮纹病抗病性21.34mm，短枝型0.99；
按照上述方案所述的方法得到待测苹果属植物样本的基因型；根据所述待测苹果属植物样本的基因型，按照所述的群体平均值和上述方案所述的基因型效应值或基因型组合效应值，利用如下标准判定待测苹果属植物样本的性状表型或者使用如下预测模型计算待测苹果属植物样本的性状的预测表型值：
(1)炭疽叶枯病抗病性：
当S1202的基因型为CC且zhwy64的基因型为CC时，判定为抗病；其它基因型判定为感病；
(2)果形：
当newdy202的基因型为CC、SIZE2270的基因型为GG、SIZE5253的基因型为CC、SIZE9100的基因型为GG或者SIZE9195的基因型为AA时，判定果形为圆锥-圆；
当SP031的基因型为CC、SP081的基因型不为CT或XDY231的基因型为GG时，判定果形为扁圆-圆；
(3)绿原酸含量或原花青素B2含量采用基因型组合模型；
按照绿原酸的分子标记的基因型或原花青素B2含量的分子标记的基因型组合估算效应值，用基因型组合效应值建立GAP模型，公式为：
GPV＝α×(GcE+μ)+β；
GPV为预测的表型值；GcE为该性状各个标记的基因型组合效应值；μ为训练群体该性状的表型均值；α和β分别为线性回归系数和剩余参数；
(4)果实成熟期、可溶性固形物含量、果汁pH值、采收时果肉硬度、采收时果肉脆度、果肉硬度保持性、果肉脆度保持性或果实轮纹病抗病性采用加性模型，公式为：
k
GPV＝α×(∑GE+μ)+β
i＝1
GPV为预测的表型值；GE为该性状标记的基因型效应值；k为该性状的标记数；μ为训练群体该性状的表型均值；α和β分别为线性回归系数和剩余参数；
(6)平均单果重、苹果酸含量、果面着色度和短枝型采用固定效应模型，预测公式为：
GPV为预测的表型值；Fx为固定效应标记基因型效应值；GnE为该性状非固定效应标记的基因型效应值；k为该性状的非固定效应的标记数；μ为训练群体该性状的表型均值；γ为缩值系数；α和β分别为线性回归系数和剩余参数；
平均单果重的固定效应为：XDY160的基因型为AA或SIZE4849的基因型为GG或SIZE4161的基因型为GG，Fx分别为-104.8，-100.7和-101.4；
苹果酸含量的固定效应为Ma、MA202和SAUR-5的基因型组合效应值；
果面着色度的固定效应分别为ZZZ162与zwy6和ZZZ162与color1245的基因型组合效应值；
短枝型的固定效应为neww45与S1245和neww45与ww19的基因型组合效应值；
本发明还提供了上述方案所述苹果属植物性状相关分子标记或者所述基因型效应值或基因型组合效应值或者所述引物组的如下一种或几种应用：
1)苹果属植物性状鉴定；2)苹果属植物指纹图谱或分子身份证构建；3)苹果属种质资源基因型鉴定；4)苹果属植物杂交育种；5)苹果属植物新品种分子DUS测试；
所述苹果属植物杂交育种包括：杂交亲本的选择与杂交组合的选配、杂交世代数设计和杂种后代分子辅助选择中的一种或几种。
下面将结合本发明中的实施例，对本发明中的技术方案进行清楚、完整地描述。显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
实施例1
1.全基因组QTL位点的挖掘。
为使所得QTL更具有通用性，试验材料选用‘红玉’ב金冠’，‘紫塞明珠’ב红富士’，‘紫塞明珠’ב金冠’和‘新红星’ב宫崎短枝富士’等4个杂种后代分离群体，杂交后代单株数分别有1568株，1679株，2211株和2019株。其中‘红玉’和‘金冠’为现代苹果(Malusdomestica Borkh.)的7个始祖品种(Founder cultivars)之一，‘新红星’为另1个始祖品种‘元帅’的芽变品种，而‘红富士’则是另外2个始祖品种‘国光’和‘元帅’的直接后代。‘紫塞明珠’属于沙果(M.asiatica Nakai)，是中国古老苹果品种的代表。于2014～2020年每年对上述杂交后代单株逐株进行表型测定。测定的性状包括果实成熟期，果形，单果重，果面着色度，果实绿原酸含量，果实原花青素B2含量，果实可溶性固形物含量，果汁pH，果实苹果酸含量，采收期果肉硬度，采收期果肉脆度，果肉硬度保持，果肉脆度保持，炭疽叶枯病抗病性，果实轮纹病抗病性，短枝型等。
利用上述杂交群体单株至少3年的表型数据，采用MapQTL和BSA-seq等方法进行QTL定位。共获得上述果实品质和抗病性性状的QTL位点459个(表4)。
表4苹果果实品质和抗病性性状QTL统计表
| 序号 | 性状 | 杂交组合 | QTL总数 |
| 1 | 单果重(Fruit size) | 3 | 72 |
| 2 | 果形(Fruit shape) | 3 | 144 |
| 3 | 着色度(Degree of fruit coloration) | 3 | 10 |
| 4 | 成熟期(Fruit ripening data) | 3 | 54 |
| 5 | 可溶性固形物含量(Soluble solid) | 3 | 78 |
| 6 | 含糖量(Sugar content) | 3 | 5 |
| 7 | 含酸量(Fruit acidity) | 3 | 6 |
| 8 | 绿原酸含量(Chlorogenate) | 1 | 3 |
| 9 | 原花青素B2(Procyanidin B2) | 1 | 3 |
| 10 | 果肉脆度保持(Flesh crispness retainability) | 1 | 24 |
| 11 | 果肉硬度保持(Flesh firmness retainability) | 1 | 28 |
| 12 | 抗炭疽叶枯病(Resistance to Glomerella) | 1 | 1 |
| 13 | 抗果实轮纹病(Resistance to Botryosphaeria) | 1 | 17 |
| 14 | 短枝型(Spur tree architecture) | 1 | 14 |
| 合计 | 4 | 459 |
2.基于QTL的SNP标记开发。
上述459个QTL中有许多重合区间，去除重合区间，在QTL峰值附近预测候选基因。利用候选基因编码区、上游调控序列，或者基因间区的变异位点开发SNP或InDel标记。共筛选出318个SNP标记和1个InDel标记(表1)。该319个标记分别设计PCR引物，并用‘红玉’、‘金冠’、‘红富士’和‘紫塞明珠’等4个亲本以及3个杂交组合各6～8株杂种后代验证PCR引物的扩增效果和各个标记在杂种后代中的实际分离情况。从实际分离情况来看，各个杂交组合的杂种后代基因型呈现孟德尔分离比。证明PCR引物设计成功。该319个标记的PCR引物序列如表3所示。
3.多重PCR扩增体系的设计与AppleGAP v2.0液相芯片的开发
提取小样测试样本基因组DNA，用
或荧光定量PCR对基因组进行精确定量。采用0.2ml PCR管/96孔PCR板，配置30μL反应体系：引物组合8μL、MP004_Cu Panel Mix 8μL、DNA 50～200ng、3xT酶10μL和余量的H2O；所述引物组合中每条引物浓度为0.24μM。按如下程序进行PCR扩增：热启动，95℃3min；16个循环，95℃30s，60℃4min；延伸72℃4min；保温10℃。
然后用AMPureXPBeads纯化试剂盒对PCR扩增产物进行纯化。用PCR扩增产物的回收物进行Illumina测序文库构建。在上述带有磁珠的PCR产物纯化管中配置30μL反应体系：3XM酶10μL；PCR primer F 1μL；Barcode XXR(10μM)1μL；H2O 18μL。按如下程序进行PCR扩增：热启动，95℃3min；6～8个循环，95℃15s，58℃15s，72℃30s；延伸72℃4min；保温10℃。最后再用AMPureXPBeads纯化试剂盒对PCR扩增产物进行纯化。
上述纯化后的PCR产物进行DNA浓度测定，链接不同Barcode的PCR产物等量混合即为AppleGAP v2.0液相芯片，直接上机测序。利用Illumina X10测序平台，采用PE150策略进行测序，测序深度为1000×～1200×。小样测试结果与前期试验KASP分型相符，表明AppleGAP v2.0开发成功。
4.训练群体的构建与基因分型
分别从4个杂交组合(‘红玉’ב金冠’，‘紫塞明珠’ב富士’，‘紫塞明珠’ב金冠’和‘新红星’ב宫崎短枝富士’)中随机机选取有3年以上性状表型数据的杂种后代各350株，再加上536份有至少3年表型数据的苹果属种质资源组成的训练群体，共1936个单株。具体表型性状为果实成熟期、平均单果重、果形、果面着色度、果实可溶性固形物含量、果汁pH值、苹果酸含量、绿原酸含量、原花青素B2含量、采收时果肉硬度、采收时果肉脆度、果肉硬度保持性、果肉脆度保持性、果实轮纹病抗病性、炭疽叶枯病抗病性和短枝型等，共计16个性状。采该1936个单株的叶片样本，提取基因组DNA，用AppleGAP v2.0进行基因型分析，获得319标记1936个单株的基因型数据。
5.标记基因型效应值的估算
利用上述训练群体的1936个单株的319标记的基因型数据和16个性状的表型数据估算标记基因型效应值，公式为：
GE(marker genotype effect)为某标记的某基因型的效应值
P(individual phenotype value)为该标记该基因型的某个体的表型值
m为训练群体该标记该基因型的个体数
μ为训练群体该性状的表型均值
利用上述公式，估算出上述16个性状的上述319个标记的基因型效应值，如表2所示。
6.GAP模型建立与检验
上述16个性状分别适用于5种GAP预测模型：
上述GAP模型经模拟选择检验，选择准确性、选择效率和选择穷进度如表5。
表5苹果基因组辅助预测芯片AppleGAP v2.0技术参数表
| 性状 | 单位 | 选择法 | 标记数 | 训练群体样本数 | 准确性* | 选择效率** | 穷尽度*** | |
| 1 | 果实成熟期 | DAFB | GAP | 52 | 1335 | 0.62 | 77.6 | 39.9 |
| 2 | 果形 | MAS | 23 | 1339 | - | 34.1 | 77.2 | |
| 3 | 平均单果重 | g | GAP | 72 | 1450 | 0.85 | 66.1 | 68.7 |
| 4 | 果面着色度 | ％ | GAP | 10 | 1327 | 0.56 | 65.5 | 67.9 |
| 5 | 可溶性固形物含量 | ％ | GAP | 76 | 1435 | 0.40 | 62.7 | 59.0 |
| 6 | 果汁pH | GAP | 5 | 1326 | 0.64 | 71.8 | 72.1 | |
| 7 | 苹果酸含量 | mg/mL | GAP | 5 | 201 | 0.66 | 82.2 | 82.9 |
| 8 | 绿原酸含量 | mg/kg | GAP | 3 | 274 | 0.50 | 85.5 | 65.3 |
| 9 | 原花青素B2含量 | mg/kg | GAP | 3 | 301 | 0.95 | 65.9 | 98.2 |
| 10 | 采收时果肉硬度 | kg/cm2 | GAP | 46 | 1272 | 0.55 | 81.8 | 98.3 |
| 11 | 采收时果肉脆度 | kg/cm2 | GAP | 46 | 1272 | 0.29 | 62.9 | 84.0 |
| 12 | 果肉硬度保持性 | 月 | GAP | 46 | 917 | 0.49 | 65.4 | 42.9 |
| 13 | 果肉脆度保持性 | 月 | GAP | 46 | 918 | 0.40 | 62.1 | 24.0 |
| 14 | 果实轮纹病抗病性 | GAP | 18 | 399 | 0.32 | 65.7 | 43.2 | |
| 15 | 炭疽叶枯病抗病性 | MAS | 2 | 780 | - | 96.0 | 94.5 | |
| 16 | 短枝型 | GAP | 13 | 487 | 0.86 | 72.7 | 34.8 |
*准确性：预测表型值与实测表型值之间的线性相关系数。
**选择效率：GAP入选个体中预测表型值与实测表型值相符的个体所占的百分率。
***穷尽度：GAP入选个体中预测表型值与实测表型值相符的个体占待测群体中符合选择标准的个体总数的百分率。
实施例2：杂种后代选择
2020年3月，采用上述AppleGAP v2.0芯片对2016～2018年配置的13个苹果杂交组合的16,214株杂种后代进行基因组辅助预测。该批杂种后代定植于河北省秦皇岛市北戴河新区前朱建坨村，为2～4年生自根实生苗，栽植密度为0.6×0.2。取样AppleGAP v2.0检测所需的叶片样品时，实生苗高度为2.0m。以0.5cm打孔器打6片叶盘置于96孔板中，做好标记，整板取满后将96孔板放入预装10g蓝色硅胶的塑料袋中扎口密封。3日后叶盘自然干燥，将96孔板从塑料袋中取出，加乳胶软盖，送交公司做DNA提取、芯片检测和基因分型。获得所有标记的基因型数据后，将基因型数据带入各个性状的GAP模型，系统自动计算出GPV值。依据GPV值对下列性状进行辅助选择。选择标准分别为：果实成熟期为晚熟(170～185DAFB)、平均单果重100g～250g，果面着色度70％以上、果实含糖量14.5％以上、苹果酸含量3.0～10.0mg/mL、绿原酸含量低于1.0mg/g、冷藏果肉硬度和脆度均保持5个月以上、抗炭疽叶枯病。按照上述选择标准，从16,214株实生苗中中筛选出优良单株77株，入选率0.475％，理论选择效率9.38％，预期选育出符合上述选择标准的新品种7个(表6)。
表6 AppleGAP v2.0选择苹果杂交后代实际效果表
实施例3：杂种后代选择
2020年5月，采用上述AppleGAP v2.0芯片对2019年配置的4个苹果杂交组合的3,404株杂种后代进行基因组辅助预测，AppleGAP v2.0检测方法和性状选择标准同实施例1。该批杂种实生苗为2020年3月播种于32孔穴盘，取AppleGAP v2.0芯片检测叶片样品时期为2020年4月，彼时实生苗为4片真叶。取其中1片真叶，取样方法同实施例1。按此标准从3,404株实生苗中筛选出优良单株11株，入选率为0.323％，理论选择效率仍为9.38％，预期选育出符合上述选择标准的新品种1个。
实施例4：杂交亲本选择
2021年2月，采用AppleGAP v2.0辅助选择杂交亲本。苹果新品种‘中农101’为本实验室杂交选育的晚熟、抗病、耐贮藏新品种，亲本组合为‘紫塞明珠’ב红富士’。‘中农101’成熟期为10月下旬，苹果腐烂病、苹果轮纹病和早期落叶病等主要病害的田间发病率均明显低于‘红富士’等主栽品种，但该品种平均单果重97g，果个偏小，果肉硬而不脆。计划以‘中农101’为亲本之一，配置杂交组合，选育大果、红色、耐贮藏、抗炭疽叶枯病新品种。
(1)侧重大果型和红色。利用Apple GAP v2.0对苹果种质资源进行所有319个标记进行基因型检测，通过基因型匹配，选配最优杂交组合为‘中农101’ב66-014’。‘66-014’为‘红津轻’ב红富士’的杂种后代优系。该2品种着色度主基因标记均为杂合基因型，因此杂种后代全红果实单株的入选率为1/4。双亲均抗炭疽叶枯病，故杂种后代全都抗病。‘中农101’平均单果重主效标记S2987和XDY160基因型均为杂合，而‘66-014’所有平均单果重的主效标记均为大果纯合基因型，因此，杂种后代大果型入选率为1/4。果肉脆度保持性状的4个主效标记在杂种后代均为1:1分离，因而果肉脆度耐贮藏入选率为1/16。综上，该杂交组合入选率为1/256，杂种群体规模应大于256株。该杂交组合于2021年4月21～24日实施田间授粉杂交。
(2)侧重耐贮藏和果面红色。经Apple GAP v2.0基因型匹配，选配最优杂交组合为‘中农101’ב17-199’。‘17-199’为‘中农101’的全同胞姊妹系。该2品种着色度主基因标记均为杂合基因型，因此杂种后代全红果实单株的入选率为<1/4。双亲均抗炭疽叶枯病，故杂种后代全都抗病。‘中农101’平均单果重主效标记S2987和XDY160基因型均为杂合，而17-199’平均单果重的主效标记S2987、S4161和XDY160均为杂合基因型，因此，杂种后代大果型入选率为1/32。果肉脆度保持性状的4个主效标记中有3个在杂种后代均为1:1分离，因而果肉脆度耐贮藏入选率为1/8。综上，该杂交组合入选率为1/1024，杂种群体规模应大于1024株。该杂交组合于2021年4月21～24日实施田间授粉杂交。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 中国农业大学
<120> 一种苹果优质抗病基因组辅助预测方法及其应用
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attaatgcca tgcacataat ttcac 25
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caaatagcta acaagaacag tggt 24
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gagcccaaac tgttattgca aa 22
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attcctcaat tggtttgtct gcgtt 25
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tcagggggtc aaacgacaat t 21
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agacagcaaa cgactaaaag ag 22
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ctctctcctc tatctcaaac ccaaag 26
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attttccaag tcgtctcgtt t 21
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tttcctggat ctgctggatt tt 22
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cgaggtaaga ggatatactt g 21
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gtggatgagg ccatgactct aagagaa 27
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ctcccaaaag ttagggcaag ataagg 26
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cttgactttt tccttatcgt ctt 23
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ggaagcggct ctgaagacta gtattagg 28
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caaacgactt tagtctttca atagttgagt t 31
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acatcatgga gagtatttcc tcc 23
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ctaatatagc tggatttaat gt 22
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accaaatgga tgaattttgg agt 23
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ccgaacggag aaattcttca taagg 25
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tgaaatcatg ctatatttag atgtacttt 29
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aactactaac tgaggagaat gtgcgc 26
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gcttcagttc atagaggaga cgaccaggaa c 31
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aaactcagag atgggatgag a 21
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gaagaagtac gagcttgtca taacaac 27
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caacaacact gaggcattgt caat 24
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gcataagtga tttaatccac cctc 24
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cattggaaat gtggtgctta tgagtaat 28
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gaagatgagg catgcttaaa agattcc 27
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tgggggaccg ataagtcttt agattaat 28
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tttcgatcat gaaggttagg aagg 24
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cgaaagcaga ggcactttat atcc 24
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attcacgctt catctctttc ctaagatagg 30
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gaaaaggggc atgcaaaaat cattttcctt 30
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aatcttgagt gatgggaaga gg 22
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gataaagggc tgtagatcct tcca 24
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cgtttgttga agctgagtat aggggt 26
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catgctttgt tcgatcatgt tgtaccaaa 29
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tgatgaaatg agaggaacat ggctg 25
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tcgaacttgt ccctacagtt cagagca 27
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gccaaattta aaatttagca agttttgggc tcg 33
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<400> 90
aaaggcactt acaaacaagt cataaaacaa atg 33
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ccgtacattt cccagttatt ag 22
<210> 92
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 92
tgtccagtct ctctgtaata ctctggaa 28
<210> 93
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 93
tagattaagg agctgacaga gttcg 25
<210> 94
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 94
agtagttttc aacctatcca tcaccggtt 29
<210> 95
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 95
tgaaaagatt caccaaatcc cagcc 25
<210> 96
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 96
gctcagtttg ctcttggagt acttgatggc gc 32
<210> 97
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 97
ttctcccata ctgatcatgt gact 24
<210> 98
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 98
attgccaaag ctattattgg taatga 26
<210> 99
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 99
gctgaaattc taactctgtt tgacatc 27
<210> 100
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 100
cagtgtcaag ctatggcaaa tg 22
<210> 101
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 101
aattggttca aaatcctcaa aggcctt 27
<210> 102
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 102
tttttctttg ataaaatcta tggcagg 27
<210> 103
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 103
ccgaggtgct acttgattgt tgcaaa 26
<210> 104
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 104
ttgtaccaat gtcctacaac cgtgg 25
<210> 105
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 105
aagaagtaga gggagagaga gagcca 26
<210> 106
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 106
ttttctgggt gttttaatgc g 21
<210> 107
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 107
attacgttct tgatatgcgg attt 24
<210> 108
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 108
gcaagatgag ttttaatgaa ccc 23
<210> 109
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 109
gaagcatcat tatcacacga atcag 25
<210> 110
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 110
atatagaacc acaggaggaa tgttg 25
<210> 111
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 111
ataaatatta aatagtctaa ggggtg 26
<210> 112
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 112
ccctcactcg ttcccttctt ttcttttctt t 31
<210> 113
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 113
aatgatgttt gtcttcataa tcaccccca 29
<210> 114
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 114
atcgtgactt gggaaacaat tctttc 26
<210> 115
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 115
aaaacataac gaggtgatta tggggtcgta 30
<210> 116
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 116
tgacatagtg gcaaataaga ggtt 24
<210> 117
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 117
ctagaacgtc tagaaaacca ggatcc 26
<210> 118
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 118
agatcgacaa gtattcttcg cttacaact 29
<210> 119
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 119
tccaaaattt gcttcataag acacagt 27
<210> 120
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 120
tttcgacatt tggatttgaa agttta 26
<210> 121
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 121
ttcaaacaga ggtaacctga g 21
<210> 122
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 122
agtagaggtg tttaaggtga gatgc 25
<210> 123
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 123
tctttccatt ggtttgacac tgatagattg 30
<210> 124
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 124
ctataacgcc atttcttctt ctgtgttggt 30
<210> 125
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 125
tctttgacga gaatcgaatc taagacctc 29
<210> 126
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 126
catgggtaat gttttgtttt gagtgc 26
<210> 127
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 127
ttaaacccaa gaccaagcat aagtct 26
<210> 128
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 128
ctcatagctg catatttcgt cgttg 25
<210> 129
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 129
acctcaaaat ttgtctggtt ga 22
<210> 130
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 130
aaatcccaga tcagaatttt tcccaaaaaa 30
<210> 131
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 131
aatacaccgc tcttgaataa tttcccc 27
<210> 132
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 132
agatcactct accaggtacg acaaatttcc aa 32
<210> 133
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 133
aagggtcaat ttgttaaatc ttatcatgac a 31
<210> 134
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 134
gcgttaatta tttgttgctt atttcaattt 30
<210> 135
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 135
tttgatgacg atgaaaactt ctgggatc 28
<210> 136
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 136
ttggatcaaa ctaagagaat gcagcaaa 28
<210> 137
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 137
tttgaaaact gctgtcagat tcttgtga 28
<210> 138
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 138
gaagttgaac atggtgtggt ttgaaa 26
<210> 139
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 139
tagaggaaac catcttcgtc tttgttttc 29
<210> 140
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 140
aggtatctgg ttcctgctga tttgactgtt g 31
<210> 141
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 141
tgccattttc agagtgcaat atgcta 26
<210> 142
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 142
tccatagcat tattacggtt tgtgcttga 29
<210> 143
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 143
aaaatgagga tatattcacg caca 24
<210> 144
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 144
ctcatcccag atatgacatg gcat 24
<210> 145
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 145
tggccacaaa ggaaaaatat ccacata 27
<210> 146
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 146
cctccagcat ggttattaac aaga 24
<210> 147
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 147
gaatctggac caacactgat agtcaa 26
<210> 148
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 148
aggctctgaa taatctcgtt t 21
<210> 149
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 149
gcgatatgtt gacaatttct ccaat 25
<210> 150
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 150
ggtgcacata tcaaagaatc tgctaca 27
<210> 151
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 151
gtgtatactc tatgtgtaat ttctcctta 29
<210> 152
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 152
taagaataat cctatgttgc a 21
<210> 153
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 153
tggaagtttg tcgatatgcc tttgt 25
<210> 154
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 154
caacgtgtgt gatatttgag tct 23
<210> 155
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 155
ctcgttggat gtcggattct g 21
<210> 156
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 156
gctttaacac tctctcctcg ctata 25
<210> 157
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 157
ctttctgggt aggaataaca cgatttgtt 29
<210> 158
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 158
gagaccaaac tcttttctct tgt 23
<210> 159
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 159
tttataagac gaagacccct tga 23
<210> 160
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 160
aaagtcaaca tcttgttcgc g 21
<210> 161
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 161
gttccagtga catcaagcat aagtctc 27
<210> 162
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 162
aaattgaagg atgtccatct ttac 24
<210> 163
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 163
tttgggaatt attcgttgtt tcgca 25
<210> 164
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 164
caccgtactt ggttatgtcc agtat 25
<210> 165
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 165
ttgatcctct actctcgaag accaagatca tc 32
<210> 166
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 166
aaagttgaaa gaggatgctg gccc 24
<210> 167
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 167
cattctgttg ggtttccgc 19
<210> 168
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 168
gagaccgact caacaatggt agt 23
<210> 169
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 169
gttatgtgat gtattatttc acgtgt 26
<210> 170
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 170
ctaacacact tggtattatt atttcagt 28
<210> 171
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 171
aattctctgt ttgattgcca t 21
<210> 172
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 172
tttttgtagg catttgatcg ggtt 24
<210> 173
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 173
tcgtaagcaa cacggcgaaa agatat 26
<210> 174
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 174
agaagctgat gaatctcagt gtttctga 28
<210> 175
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 175
tccttcggaa agagagagaa acgag 25
<210> 176
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 176
atcgaataaa taccaactcc cacttggaat 30
<210> 177
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 177
atgaccaata aatctagaac aagagtagca 30
<210> 178
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 178
atctgcactg tacacgtgta tacatc 26
<210> 179
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 179
tgcacatttc cactaaccga atatgtat 28
<210> 180
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 180
aattaaacta ctgtatgtgc gcct 24
<210> 181
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 181
ccagtcatac tgactgtaga ggaacctcgt 30
<210> 182
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 182
ttgtaaattt caattctcac c 21
<210> 183
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 183
tcaaggcacg attcctaatt ggattt 26
<210> 184
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 184
ccctgaagtt gatttgagtg aaggt 25
<210> 185
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 185
gccaaattta tgccacatgg caa 23
<210> 186
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 186
ggttcgtaca gaaatgggag ca 22
<210> 187
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 187
atgtttatgg cattgctgat aca 23
<210> 188
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 188
gtcgaagtga acatgtaata gtgcca 26
<210> 189
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 189
gagtcacacc taccgaactg ttaggtag 28
<210> 190
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 190
ttgaagtaag ggtagattgt gt 22
<210> 191
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 191
atttccataa acttcacagg caatga 26
<210> 192
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 192
cttctaaacc cggcaaacta agttctattg 30
<210> 193
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 193
taaccatgtt gaaattctcg ccctaca 27
<210> 194
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 194
gatgtaaccc ttcttgagct gttca 25
<210> 195
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 195
aacgagatat cgacatgatt tgt 23
<210> 196
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 196
catgcatcaa gaagaagctc a 21
<210> 197
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 197
tgatgatgtc aaaacaacac gcttaac 27
<210> 198
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 198
aaatgttgca ttctgtagag ctcat 25
<210> 199
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 199
ctcatatcct ttcaatgtcc agc 23
<210> 200
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 200
tacataacct cctgtcctga ttgttcag 28
<210> 201
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 201
caagagcaat taacgccatc ctaattt 27
<210> 202
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 202
cgagtggtgt ttttgtttga ttcttg 26
<210> 203
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 203
aaatggccac atcttcaatt tgca 24
<210> 204
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 204
tccagacata ttttgagtgt cacttgtca 29
<210> 205
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 205
atgcagacat aaggggaaca ag 22
<210> 206
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 206
ccagaataca gtgggaactt ctcaacg 27
<210> 207
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 207
tggtttgctc actaaattgg atgg 24
<210> 208
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 208
gcacaaatca ttttcgcgaa gatgacaca 29
<210> 209
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 209
atcacaacat atccaatttt gcattt 26
<210> 210
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 210
ggatggttct tgtcatttaa gct 23
<210> 211
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 211
aaaacagttc gaaggttggt ct 22
<210> 212
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 212
agtcttggaa acgaagtaca tct 23
<210> 213
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 213
cagtttcaat ataccaaagg cagcagta 28
<210> 214
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 214
cttttgtgaa tccaagtaaa tgggg 25
<210> 215
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 215
ggaatacaac ttttccaacc ccaagtgcac 30
<210> 216
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 216
cacccttttc ctttatgctt catagaac 28
<210> 217
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 217
cagcaaactc atgttcttca agtagctct 29
<210> 218
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 218
tggagctgtc ttgtggccta attaatcac 29
<210> 219
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 219
ctcgttaaac tttgttggat aactct 26
<210> 220
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 220
ttttggtcaa agtcatccgt ttgcttacgc 30
<210> 221
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 221
atggttgcat atgaagaagc aactct 26
<210> 222
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 222
caagttcttc aacatatcaa gccca 25
<210> 223
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 223
cttggaaatc ccagttgagg ttgaaggg 28
<210> 224
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 224
ttatatagct tagagggctt tcttacgggc t 31
<210> 225
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 225
accccatttt tgttgtggac c 21
<210> 226
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 226
ggtgaggtta gtgtggtcga attctaat 28
<210> 227
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 227
agtgatgatg ttcctaagat tgt 23
<210> 228
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 228
ggcttagagt ataccaaaat accg 24
<210> 229
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 229
ccaatgtggg atgaatcaag aagcatt 27
<210> 230
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 230
tcaatcacag ttcgtggctg ttc 23
<210> 231
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 231
gttgagcgta tgaatcctat at 22
<210> 232
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 232
atggaggact atagaaagag aaaacgaaag t 31
<210> 233
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 233
gctatgttta gggacaggaa gt 22
<210> 234
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 234
tgagatcaca ttaacatgta gc 22
<210> 235
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 235
acatgtacga ttcatgggca acc 23
<210> 236
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 236
gtgcaaaggt gaacacagga caaca 25
<210> 237
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 237
cgagtcagtt taggacccct atat 24
<210> 238
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 238
catcatgtcc acaagaaccc taa 23
<210> 239
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 239
cttggtatta tttactttac gttttatttt gtctttat 38
<210> 240
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 240
tttacaacca aaatcattgc agatgcggt 29
<210> 241
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 241
cactcgctgc atcttaaact attatga 27
<210> 242
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 242
ttgtgagggg atcatatctg tagtcgt 27
<210> 243
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 243
actacacaaa tgggatgaga ctttcca 27
<210> 244
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 244
tacagagacc ttcttctcat aacca 25
<210> 245
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 245
gagttcgcgt attctacaat tat 23
<210> 246
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 246
aaagtcttta catctccgca tgaatc 26
<210> 247
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 247
ttcctaagaa accctagctt ccattt 26
<210> 248
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 248
gatttgttca gaccagtatg ttcctt 26
<210> 249
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 249
caaaatttct ccttcaaact ca 22
<210> 250
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 250
atgtgcctac aagaaattaa atgggt 26
<210> 251
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 251
gcgttgtctt ccatgatttt cccaagt 27
<210> 252
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 252
aattttggcc agaatacgaa caa 23
<210> 253
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 253
tcgcttttag ccaacactaa ttatt 25
<210> 254
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 254
agaggaagaa gaaaacatgg ggta 24
<210> 255
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 255
aatagctaga caatttaatg agtttgt 27
<210> 256
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 256
aattccatct caagtgtcag ttgccaa 27
<210> 257
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 257
atgagtgagc gtttggtttt ccgt 24
<210> 258
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 258
ccgcagcatc aatcataatt atatgcaact 30
<210> 259
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 259
acaaatatct ctcgcatgtc tttgtaatt 29
<210> 260
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 260
gttctctctt taatttcctg gtgttgc 27
<210> 261
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 261
aggttccaaa tttaaatctc cctcgca 27
<210> 262
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 262
gcgcagtgta agaggaaaat gttga 25
<210> 263
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 263
aagaccaact agagtggata gctaacttta a 31
<210> 264
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 264
tccataaaaa ggacacaaga cctg 24
<210> 265
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 265
gtgggctcat ataaacaaca aataag 26
<210> 266
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 266
accactaaat gtttgatcct tgtc 24
<210> 267
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 267
atctgctaca gagaaaaacg gtaaaccatc t 31
<210> 268
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 268
cacttgtgca attcaaaatt tc 22
<210> 269
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 269
aattatcttg ccgagctttt tgtcct 26
<210> 270
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 270
ccctagctcc tttcgaccct tttaatgacc 30
<210> 271
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 271
tctgttctgt ctcctagttt gtatgg 26
<210> 272
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 272
agagagtgaa ggtagctcca tc 22
<210> 273
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 273
gtttctagtg tatgggccaa ctc 23
<210> 274
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 274
acgataatgt aactcagctc a 21
<210> 275
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 275
taggaggttc aaatttgaga tatttttcaa c 31
<210> 276
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 276
tcatgagagt cttgtatagg gttggt 26
<210> 277
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 277
ctaaaaacca taccagtgaa ccaacc 26
<210> 278
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 278
aattttttca caccgtttga actcga 26
<210> 279
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 279
tgcagacaca acagtaagtt ctcc 24
<210> 280
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 280
ctggaatgtg taccaaaatc tcaaata 27
<210> 281
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 281
tttcaccaat ggtcttatat tcagaaagac ta 32
<210> 282
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 282
gagaattccc ctattttgaa 20
<210> 283
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 283
caaccaccat ccgtgaacat ataaac 26
<210> 284
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 284
aaagggaccc aatattttgt ttgtttgt 28
<210> 285
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 285
caacagaatg tccgttagaa ctcagatt 28
<210> 286
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 286
atgagtatgc agacgggaaa gaactag 27
<210> 287
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 287
gtgagagtga tggtcaagtg ataaga 26
<210> 288
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 288
tgctacattc ccccaaattt agtt 24
<210> 289
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 289
caaccaaatc aatgaacaag ggaagaata 29
<210> 290
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 290
aacgtgatca tgcatctgtt gaag 24
<210> 291
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 291
tgaaaaaaca tgaaagtcgc atgaatgt 28
<210> 292
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 292
gcgccttata caagtgttat atggtg 26
<210> 293
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 293
tgaagagacg taaggatgcc tcac 24
<210> 294
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 294
ctcaagtgtt gcgaagatga tacag 25
<210> 295
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 295
aacgtaaatt tgtttggttt gag 23
<210> 296
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 296
gaacaataac ttgcctgagg aaaagtctcc a 31
<210> 297
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 297
cctcatggag attcatttcc cttcct 26
<210> 298
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 298
aacgagctgg tgtaagtttc ctataaatgg 30
<210> 299
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 299
ctgagagact tggccaagaa atacgg 26
<210> 300
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 300
ggtgcaaagc caatgtttgt atcacc 26
<210> 301
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 301
agaagatacg cggtttgttt t 21
<210> 302
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 302
aaaagtcacc ccttaaattg gaagaggcg 29
<210> 303
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 303
acaagactac catagacccc cttatgcttc 30
<210> 304
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 304
acttgaaagg cgggaaattg tatttgag 28
<210> 305
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 305
caaacgtgaa aatctgagga taagttccct g 31
<210> 306
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 306
tggggttgtt ctaatgtaat ctgcagc 27
<210> 307
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 307
cactgcgtgc atatacatac attcatta 28
<210> 308
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 308
tattatttct tagattcaca ggtttcg 27
<210> 309
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 309
caaggagttc ctttcattct catccaaccc 30
<210> 310
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 310
actgaagtgt tgctgaagaa tgag 24
<210> 311
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 311
ggggtgttca cataggattt gtatg 25
<210> 312
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 312
agaacaacct ctttcatggc a 21
<210> 313
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 313
agtgatactt gccatgtagg actctc 26
<210> 314
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 314
tgtgtccaac gagattaggt gtg 23
<210> 315
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 315
gcatctagtc cattcgtaat gtaggcgcca a 31
<210> 316
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 316
gaggtcttat catcctgtcc atctccaga 29
<210> 317
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 317
gcaaatcttt gtattctgct tgcag 25
<210> 318
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 318
cttacccttg gtttgtactg tt 22
<210> 319
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 319
agttgtatca aagtcacgaa tca 23
<210> 320
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 320
aacacatgga aagagattga tcacctt 27
<210> 321
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 321
gtaataaaag gaggaagtgg ctgcg 25
<210> 322
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 322
cattaaatgg gtggatgagg atgaccccg 29
<210> 323
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 323
tgtgaccaga ccaaacggaa aaactctaga t 31
<210> 324
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 324
atgggtcttg aaatattaat gagattccgt tt 32
<210> 325
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 325
cggcctattt ttgctaatgg tatgct 26
<210> 326
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 326
gaaagtatga aatggaacca gtctttaa 28
<210> 327
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 327
gatagatgga agggggaaaa 20
<210> 328
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 328
agggtgggta tattagcact ctcaatta 28
<210> 329
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 329
tgacaccaac cattcaaaag tgctattg 28
<210> 330
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 330
ttgcaaatgg agagtggtgt agtgt 25
<210> 331
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 331
gatgtaagca cagtttttgt cgatcca 27
<210> 332
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 332
ggcttctttt tatcaccgag ttgaaaacg 29
<210> 333
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 333
aaaatgacag actcgattgc agc 23
<210> 334
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 334
agtagtgtta ctggctctgt atcttgttaa 30
<210> 335
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 335
aaccaagaaa caagtcatag ct 22
<210> 336
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 336
attcagttga catctgcgaa agga 24
<210> 337
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 337
cgatttctcg actggttaat cgatcatcg 29
<210> 338
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 338
aaatccagat ctgaaagcac gagatg 26
<210> 339
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 339
accagtgtaa acccctggga ttctg 25
<210> 340
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 340
tctcttcctt ctgctttctc ac 22
<210> 341
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 341
ggtagaaagt ttcgatcatg gttgg 25
<210> 342
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 342
tcacacggga acttttctca tct 23
<210> 343
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 343
cacgatcaga aaagctatct cgtctaaca 29
<210> 344
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 344
aacccatctg atgataaggg tag 23
<210> 345
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 345
gacagaagtg aaacagaaac atctgcag 28
<210> 346
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 346
gcaaagttcc aaggaaatct gatatg 26
<210> 347
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 347
atgcataatc caaagctgcc tgaagtg 27
<210> 348
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 348
ccacaactcc aacaataact aaccct 26
<210> 349
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 349
gccgttgatt cctgggtagg tga 23
<210> 350
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 350
attgccgaaa gctttaaaga ctccg 25
<210> 351
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 351
taccagatac gcataacata gtggga 26
<210> 352
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 352
cttacctgaa atcagtttgc ttcatact 28
<210> 353
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 353
ccatttccgg gaaggtgatt aattgc 26
<210> 354
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 354
gtgtcacaat cttctatcga gat 23
<210> 355
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 355
atagaaacct tgacagccaa agac 24
<210> 356
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 356
gggttcaaca caaaacggtc ctatcatta 29
<210> 357
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 357
agttttgtgg acgaaattgt cttttt 26
<210> 358
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 358
ataagaaatc gtcccactct tc 22
<210> 359
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 359
aacaatttgc gatttcctgt acttgt 26
<210> 360
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 360
gaattgggat gaggagataa gaaa 24
<210> 361
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 361
taacccttgt cataatgcag tgtttc 26
<210> 362
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 362
tgttcatgtt gttgtacagc acgc 24
<210> 363
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 363
attagtaaga caccaatgac ctcaca 26
<210> 364
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 364
gaagcatttg taatcagtcc gacattc 27
<210> 365
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 365
agttggaaat cctttttcat ccccttt 27
<210> 366
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 366
tcgcacatcc catcagtttc aatctga 27
<210> 367
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 367
ctacatctga tgactacagc ttgtgcat 28
<210> 368
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 368
catgttttcc attggatctg gat 23
<210> 369
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 369
attcatctcg tgcctatcaa ctt 23
<210> 370
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 370
tgaagacaag ggagatttaa cacattaa 28
<210> 371
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 371
cgactcatcc atccgattat aa 22
<210> 372
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 372
gatccaagaa tttaacgact ccacaacca 29
<210> 373
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 373
aattgttttc ataatactcc tctc 24
<210> 374
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 374
gcttttcgag gatccacttg aattttta 28
<210> 375
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 375
ctagccaata gagaccgatg aacaattat 29
<210> 376
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 376
gaaaatttgg gagttcaacc tgtcg 25
<210> 377
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 377
caaggaatat agcaacgctt gaagaacaag ac 32
<210> 378
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 378
attggcccaa tttcgtggtg tttg 24
<210> 379
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 379
agtgggccta aaaataaagc tt 22
<210> 380
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 380
caaaggctgt aaaaagtgta caaaaatcat tta 33
<210> 381
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 381
aacaacccct atgccatgaa ct 22
<210> 382
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 382
cgaagaggaa cagcataaga gtgtaccaaa t 31
<210> 383
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 383
gtggaattgg tggagctctg cataca 26
<210> 384
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 384
cagaaaagaa ggtgggaaac ttactc 26
<210> 385
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 385
ttcggggtcg tgagaatgtt at 22
<210> 386
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 386
ggtatgatta aactgacggt tccctc 26
<210> 387
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 387
agtaggtttg gcagattgac tttgaaatcg 30
<210> 388
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 388
agtgagttgt ttttggacca a 21
<210> 389
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 389
ttgtcccgta agtcaacaga ttcaaat 27
<210> 390
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 390
gactggaaaa cacattaaac agcatta 27
<210> 391
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 391
ttcagtgtac tgccgaattg aag 23
<210> 392
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 392
gtcttaaact gaagaacacc tctgccg 27
<210> 393
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 393
gcaagttaat catgcagaag tcat 24
<210> 394
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 394
gttcgtccat aacctagctc tttcttca 28
<210> 395
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 395
tcttgttcct tggagattca aagttg 26
<210> 396
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 396
tatagacatg gagggacaat ggtgaa 26
<210> 397
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 397
tacgtcaaaa cccagtttga tt 22
<210> 398
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 398
cagaacccat acgcttcaac 20
<210> 399
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 399
aggggaagct tattttctta gggaggttg 29
<210> 400
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 400
gttaagcgtt tgaatctgat caactgttgt 30
<210> 401
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 401
ggccttctca agttcttctg ctgtgaagat t 31
<210> 402
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 402
taatgggaag ctcttggttg tatttggg 28
<210> 403
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 403
ttcttttcag ttcttaactc cacacca 27
<210> 404
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 404
acattacaca atggcttcga gaatgca 27
<210> 405
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 405
tggaagttga tggttgaatt atcat 25
<210> 406
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 406
ttaactcaaa ttgcaggatg gaggtgt 27
<210> 407
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 407
tttattagaa gcatgtatca gcgg 24
<210> 408
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 408
cttccgtttt cctttgttgt 20
<210> 409
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 409
tagatataca agattcagca tttttt 26
<210> 410
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 410
aacggctcta atgatttgat tatttgata 29
<210> 411
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 411
tattgctagc actcgagaga attg 24
<210> 412
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 412
tgaaatgaat cgcagtggac aacaatag 28
<210> 413
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 413
cccacaaagt aaagtttctc ccacaa 26
<210> 414
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 414
gaaattggag tttttcaggg tttgtgttat 30
<210> 415
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 415
attctatgtg gtgaaagatg acaagact 28
<210> 416
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 416
gtactgtaaa ggctcaatta ccttg 25
<210> 417
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 417
agttgtcctg gaagtcatca gaccag 26
<210> 418
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 418
aggtcaaatc cttcaaacca ac 22
<210> 419
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 419
ccaactacat tgacgattgg attgttgaa 29
<210> 420
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 420
ggtttttaat cgggcaaagg tatatt 26
<210> 421
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 421
gtcctggctg cgttaacatt tttaatagg 29
<210> 422
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 422
tccaattcat gttttgtggg a 21
<210> 423
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 423
ccaaacactt tagatatcac cctcaa 26
<210> 424
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 424
acaagagttt gatgcctcta cttccaacg 29
<210> 425
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 425
ctgcatggaa aagcaaaatt cctacaat 28
<210> 426
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 426
gagttggtta tgttgatgtt gagttag 27
<210> 427
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 427
tattatcttc gactcttcct ctttgaatat a 31
<210> 428
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 428
gatttcgttg taaggtaaag ggaa 24
<210> 429
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 429
acaatagcca gattattagg tccgg 25
<210> 430
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 430
catgtgtcag cttgaatttt gtcttt 26
<210> 431
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 431
aaagatccta tagcttccgg tcgca 25
<210> 432
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 432
actagtcgat ttgaggggga aaggcta 27
<210> 433
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 433
tagttttaac gtgcatgcaa tctc 24
<210> 434
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 434
caatgggatt tctcttctcc agaacaaatg 30
<210> 435
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 435
aagtgggtca cgattgaagt tcttatcag 29
<210> 436
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 436
attcttgaat ccagcaatcc aaatac 26
<210> 437
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 437
tcaacctcaa ccgattatgg ccatatag 28
<210> 438
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 438
caaagaaggg ctctcaaaga ctacaattct ag 32
<210> 439
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 439
agaaaagttt ctcgagggtt tagtgaggga 30
<210> 440
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 440
tcatctcgtt ttgatggttt tcatg 25
<210> 441
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 441
ggggtgatgt ttattgagat tgagagct 28
<210> 442
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 442
tctcatggct tcctccactc caaaa 25
<210> 443
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 443
ttaacccaaa agaagagcca ttc 23
<210> 444
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 444
gattcagaac cttccttgcc t 21
<210> 445
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 445
aacggtaatt aagccagata tttata 26
<210> 446
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 446
agagttaaaa tgtgcactgg attg 24
<210> 447
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 447
gaaatgataa catgttttgg aatggcgta 29
<210> 448
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 448
attttcaaag tttgagggag agaacg 26
<210> 449
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 449
tggactattg gggtttgttg atgttgt 27
<210> 450
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 450
cttatgcaac gactagatct gc 22
<210> 451
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 451
attattgctc tttcttcttt gctg 24
<210> 452
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 452
ccgtacctct ctaaatacgg aatggag 27
<210> 453
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 453
agaattgttt cagaggctaa caccat 26
<210> 454
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 454
tgatatgcac cattggttgt cgga 24
<210> 455
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 455
tttttgtcga agctcacaaa ttgtgt 26
<210> 456
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 456
attgtcaagc agcaaacgat aaat 24
<210> 457
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 457
aaagccaatc caactttaca tctgaa 26
<210> 458
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 458
aggtgtgaat ttgtcctagt tttcta 26
<210> 459
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 459
gtggagaact aggtcttttc ttgga 25
<210> 460
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 460
ctcttacatg ttttactctt cattga 26
<210> 461
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 461
tacaaacttt tacatgagac gctccaaata ac 32
<210> 462
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 462
atcgcaaatt acgtgatttt aagcagtcaa g 31
<210> 463
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 463
gcattaaaac caaagccttc cctgtccgaa 30
<210> 464
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 464
atggtgggga tgattaagac gtaattaaa 29
<210> 465
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 465
ggtttctctc tctcccccta tt 22
<210> 466
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 466
tgaacttacc agatcagcaa agagtt 26
<210> 467
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 467
gtttggatgc ttatggattt tggag 25
<210> 468
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 468
ttcaggggta caaaatctag atagcaatgc 30
<210> 469
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 469
gatctgaaga ccaagacaat aacaatttt 29
<210> 470
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 470
atataatgta gttgttgatg attg 24
<210> 471
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 471
ctcaacggga agaaaagggt gtttaat 27
<210> 472
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 472
acatgcgtct aatgaagctg at 22
<210> 473
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 473
tttcattcgc ctaactgagt ttggt 25
<210> 474
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 474
attgtacgtg gagtttgatg gacttt 26
<210> 475
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 475
tgtgatgatc gtgggcattt atct 24
<210> 476
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 476
aattgtctgc aactactaaa catgacctt 29
<210> 477
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 477
aatctccttt ttcctggtca tcta 24
<210> 478
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 478
cccttgtact ggattccaat gacg 24
<210> 479
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 479
cttcttcaca tagccttgtt tccat 25
<210> 480
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 480
tgaggtacgg aattaagtca tatggcacgt 30
<210> 481
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 481
agctttcttg gcaaggaata atctt 25
<210> 482
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 482
gtctgacagc ttgattttgc tatgaat 27
<210> 483
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 483
gcttgcagga gggccctacc 20
<210> 484
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 484
tcaaaccccg acgtttgcat c 21
<210> 485
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 485
gaggaagaaa aaggggagga aagcc 25
<210> 486
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 486
ttcatagcct cctgccaaat agtgc 25
<210> 487
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 487
ctctgttttg gctagtgaaa caagtaaga 29
<210> 488
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 488
atgcatggag aaaagttttc cct 23
<210> 489
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 489
ttgttcttgt ccaaaaatca cagcagct 28
<210> 490
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 490
actttgaatg gggattttga ggtgg 25
<210> 491
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 491
aacatactct aaggtacgct cc 22
<210> 492
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 492
ggatggatcg gttttggaga taa 23
<210> 493
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 493
ctccaagccc taaggatggt attgacc 27
<210> 494
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 494
aaaagtccac gtgtcatg 18
<210> 495
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 495
cattttcccg atgctttatg taatca 26
<210> 496
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 496
gacctttact gttactaacc cctgattta 29
<210> 497
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 497
ttaatattga cgccagtaga atggacca 28
<210> 498
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 498
acgccatcaa taacgggtat gattcatg 28
<210> 499
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 499
tctccctaca ctacacactt ttgttg 26
<210> 500
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 500
gaagctctcg tgtttttatc ttgcatttt 29
<210> 501
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 501
caaaacagat tagatcgatt gggt 24
<210> 502
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 502
gcataatata caaacacttc actgc 25
<210> 503
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 503
tatggcacaa gcttgtatat aatgtcacat 30
<210> 504
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 504
gctgaattca tagctcatta gtcaatac 28
<210> 505
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 505
tgcattttcc tctgtgtatg tgt 23
<210> 506
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 506
gtatattcga cgtacaaaag atcctcca 28
<210> 507
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 507
ttaattaagc cttatttgct gggtg 25
<210> 508
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 508
ttgcaaactt gggatccata tg 22
<210> 509
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 509
aaagcatcga aaacaaaatc g 21
<210> 510
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 510
aaatccacgc aatatcaaat agcgtgg 27
<210> 511
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 511
agtgattagc tagtttcatg gcttacg 27
<210> 512
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 512
ggaaggtctt gaatgtctat gggtcttt 28
<210> 513
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 513
actctcccta tgtggtcttt caaaatt 27
<210> 514
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 514
tttgttcacc aactttacac catcc 25
<210> 515
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 515
caaacaagac aaagattcca ttttcccaat cac 33
<210> 516
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 516
ggcgatggag acggaggtcc aa 22
<210> 517
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 517
tacccatttg ccaatcgttg att 23
<210> 518
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 518
aaacccagaa ggaaatttaa acaaa 25
<210> 519
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 519
ctatgaaagt gagagctttt tgt 23
<210> 520
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 520
gcttaatcga attgagtcgc taaccca 27
<210> 521
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 521
agtagcaagc ttctgtgcaa aaca 24
<210> 522
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 522
tcctctaaac atctccacct gattg 25
<210> 523
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 523
aaatgatcaa agacgtatcc acaagacaac ta 32
<210> 524
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 524
ctccaatcat cggtgtgaaa a 21
<210> 525
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 525
actgtttggg atttgtcttc cacata 26
<210> 526
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 526
agtcagggtc tgtgtatgac ttaaaat 27
<210> 527
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 527
ggcttacagc gaatttcagt aacacctgt 29
<210> 528
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 528
attgtttgaa tttcctagac gctctcgt 28
<210> 529
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 529
gtttttaaat ccaaaaaggc caca 24
<210> 530
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 530
atggggaaga gagcaatttc t 21
<210> 531
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 531
cttggtatac ctacgttctt gtat 24
<210> 532
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 532
gtattgctcc ttgatatctg tt 22
<210> 533
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 533
atgcaatgtg cttcaacatc ttctcactca gg 32
<210> 534
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 534
aacgactttc taaggttttg atgctc 26
<210> 535
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 535
atcaaattaa tgggagctga gtgatgac 28
<210> 536
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 536
cttcagctct tgaaagactc caaatg 26
<210> 537
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 537
tcaaaatcct aattgaatgc acc 23
<210> 538
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 538
ttagtggttt acactttaca ctacatgc 28
<210> 539
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 539
ccacatttga taatccaatt cgtattt 27
<210> 540
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 540
ctttcagagt tcatgcaaag gttcttca 28
<210> 541
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 541
cttttccatt atccaaacca gaccaccct 29
<210> 542
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 542
cactcatctg ttctcatacg aagcaa 26
<210> 543
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 543
ctctaagtaa agagaatcga gctcc 25
<210> 544
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 544
gaacctgttt tgtctgagga tt 22
<210> 545
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 545
caacgaaaat aaacaggtgg tgatcg 26
<210> 546
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 546
ttttgattct tcaggttcct 20
<210> 547
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 547
tttcgagcac acaaattttg tcaatcg 27
<210> 548
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 548
ttcaagggtc cctaaaaatc ggggac 26
<210> 549
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 549
ttagtaacaa attcaacgta cccctcta 28
<210> 550
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 550
tattgacacg attgaggtac gagaaga 27
<210> 551
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 551
taaaaaacca agagactgag actct 25
<210> 552
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 552
gacgaattta aaatctttca ctc 23
<210> 553
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 553
gcaactgcaa aggtgaattc atct 24
<210> 554
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 554
catcttctcc cttgttctct cccttgcc 28
<210> 555
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 555
atctgggtag aaaacgagat ccga 24
<210> 556
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 556
ttagaggtag aggaggaggt g 21
<210> 557
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 557
gattaggcca cctgagtttg aatcc 25
<210> 558
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 558
gtggaaacta gctacttcca tct 23
<210> 559
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 559
tgcaaaacca tcttctaagg gcttta 26
<210> 560
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 560
ggttgttgct ctcccttatc ttgaaatt 28
<210> 561
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 561
cccacatgct aatgtttgct ccatcaa 27
<210> 562
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 562
acccggacaa gaaaatctca gata 24
<210> 563
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 563
ttgacacaaa agatattgag gaagga 26
<210> 564
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 564
agaggtcaat aaatttgggg acttc 25
<210> 565
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 565
aatattgtag ccatttcgta ttttcggtac g 31
<210> 566
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 566
tcggatttca tctcacttca gctgttcaat a 31
<210> 567
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 567
cagagaggac ggcgaaga 18
<210> 568
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 568
tccaacttgc gccactttgt g 21
<210> 569
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 569
aaaactgata tatgcatgag acggcttct 29
<210> 570
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 570
tttggcttgt gctcctttta tagta 25
<210> 571
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 571
aaaactgata tatgcatgag acggcttct 29
<210> 572
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 572
tttggcttgt gctcctttta tagta 25
<210> 573
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 573
ggctagacaa cttaacaaac tccggcc 27
<210> 574
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 574
gcagttcatt tttcccatca aaacccatg 29
<210> 575
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 575
gatttgtttg gttttaagca ggta 24
<210> 576
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 576
tgccatattt cgtgtggtgt gttaattt 28
<210> 577
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 577
ccatgcattg cctattccct aaattc 26
<210> 578
<211> 31
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 578
attgtaattt tactccaatg caatcacata t 31
<210> 579
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 579
attgtagagt agtgccaaaa gtgcaaatca 30
<210> 580
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 580
taactgctaa tccgaaaatt aagcga 26
<210> 581
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 581
tcctgttcag aacacattgg tttttact 28
<210> 582
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 582
actagcagaa gaagaactag aaacctctcc 30
<210> 583
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 583
ggagattcat ttgcaaagag ggttg 25
<210> 584
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 584
gagatttcct tctcacactt ttgc 24
<210> 585
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 585
atacaatctt ttaaaagggt gtcctgcata 30
<210> 586
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 586
atgagactta actcagctca actcctt 27
<210> 587
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 587
ttttaccatt tgtatgcgca atgagattt 29
<210> 588
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 588
aaaccccata accataagct gcatagcca 29
<210> 589
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 589
tctggctcct tgtaaatatg ag 22
<210> 590
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 590
tgctgtgtca gaggagcaga aaaacata 28
<210> 591
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 591
ctcaatccct tgcactggct ctagc 25
<210> 592
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 592
taggaggagt gcaaactttg tca 23
<210> 593
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 593
ttgtgtttac cacctactta tgcg 24
<210> 594
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 594
ttaacgatct agtcaaccac acttcttc 28
<210> 595
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 595
ttctttcttt attcgcgcac att 23
<210> 596
<211> 32
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 596
cgagatcgaa agctcagaag aatgacagca ct 32
<210> 597
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 597
ctggttcggt gagtttggca atagag 26
<210> 598
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 598
aatccatcta tcaccgaaca cg 22
<210> 599
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 599
ttgggaaata tttgttgatg tcattag 27
<210> 600
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 600
tgcaccacca tgagaccaat tg 22
<210> 601
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 601
gaggccaaaa ccaaagagaa aaatatt 27
<210> 602
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 602
ctatggtggc gcgtggagat atcgctt 27
<210> 603
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 603
aggcgatcct ttacgtgatg gctct 25
<210> 604
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 604
ccagaaattc gaggatgaag aagcagg 27
<210> 605
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 605
gatcctttta agtacggatt ttcc 24
<210> 606
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 606
gctgtatttg ctgaccgttg atgaat 26
<210> 607
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 607
agttgcgaaa ttctacacct tctaccactg 30
<210> 608
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 608
gtagaagctt gccagtttgt cgagg 25
<210> 609
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 609
ctacttaaag cattcaggct 20
<210> 610
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 610
taaatcttag tgtatgctca ccat 24
<210> 611
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 611
catttcagta ttcctgtcgt gg 22
<210> 612
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 612
ctcccaaact tgactttcct catcag 26
<210> 613
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 613
gactgtagat accaagtgag tactgt 26
<210> 614
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 614
ctgaatcatt tgattgtttt ggacaac 27
<210> 615
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 615
tgactcctcg gataactctc actt 24
<210> 616
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 616
ttgtgtgtgg ttggtttaca tg 22
<210> 617
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 617
ctatgtttat aactagaacc gg 22
<210> 618
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 618
aactaaggtc gtttagtcaa ccaacgta 28
<210> 619
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 619
aagccccaag ttttgtgaac agacac 26
<210> 620
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 620
gtaaataccc tgccaactag gtac 24
<210> 621
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 621
ggcgatgaag aagtttgtcc ctctct 26
<210> 622
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 622
tagaacaagc acgaactttg ttttcga 27
<210> 623
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 623
gacatggaag tattttaata gtaaaatgct 30
<210> 624
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 624
agaaatccct gtgcataaag ggctatata 29
<210> 625
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 625
ctgctcctat actggcaatt ta 22
<210> 626
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 626
ttgttctttg ccatcttgtc attcat 26
<210> 627
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 627
caaggggaag gaacaaaaga tatta 25
<210> 628
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 628
tgcttgtgat gttcatcaga aatgcggtgc 30
<210> 629
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 629
ggaagttttg aagcaataag aggcctta 28
<210> 630
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 630
gccaaggcac atcaatgtat ttt 23
<210> 631
<211> 28
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 631
aggattcaat ttggattttc acccacgg 28
<210> 632
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 632
gctttgggtt ggtaattggc tact 24
<210> 633
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 633
cttctgcaaa accatattga tag 23
<210> 634
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 634
ctatatgagg atttcagaca gtctg 25
<210> 635
<211> 29
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 635
gcagcacaaa attcatcaca acaaaatcc 29
<210> 636
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 636
tgggggttat attgggggtt gtgatt 26
<210> 637
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 637
attgttctat gcttctaacc accg 24
<210> 638
<211> 35
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 638
acaaatttaa tttttctagc attacttatg tttag 35
Claims (7)
1.苹果属植物性状相关分子标记组合，所述苹果属植物性状相关分子标记组合对应的SNP位点和InDel位点如表1所示；所述分子标记组合由表3所示引物组合扩增得到；
表1苹果属植物性状相关分子标记组合对应的SNP位点和InDel位点
表3检测不同分子标记的引物组
2.检测权利要求1所述分子标记的引物组合，由SEQ ID NO.1～SEQ ID NO.638所示的引物组合组成；
所述引物组合对应检测的标记的名称、上游引物和下游引物如表3所示。
3.一种权利要求1所述分子标记的检测方法，包括以下步骤：
1)提取待测苹果属植物的基因组DNA；
2)采用权利要求2所述引物组合对待测苹果属植物的基因组DNA进行多重PCR扩增，得到扩增产物；
3)采用二代测序方法测得所述扩增产物的基因型，得到待测苹果属植物样本的基因型。
4.根据权利要求3所述的方法，其特征在于，步骤2)中所述多重PCR扩增的反应体系以30μL计，包括以下组分：权利要求2中所述引物组合8μL、MP004_Cu Panel Mix 8μL、DNA 50～200ng、3xT酶10μL和余量的H2O；所述引物组合中每条引物的浓度为0.24μM；所述多重PCR扩增的反应程序包括：95℃3min；95℃30s，60℃4min，16个循环；72℃4min延伸。
5.根据权利要求3所述的方法，其特征在于，步骤3)中所述二代测序深度为1200x。
6.一种判定苹果属植物样本的性状表型或者计算苹果属植物样本的性状的预测表型值的方法，包括以下步骤：
权利要求1所述的分子标记对应的性状的群体平均值，分别为：果实成熟期159.45DAFB，果面着色度56.35％，平均单果重106.63g，可溶性固形物含量14.85％，果汁pH3.34，苹果酸含量5.83mg/mL，采收时果肉硬度12.18kg/cm2，采收时果肉脆度1.31kg/cm2，果肉硬度保持2.41月、果肉脆度保持2.19月，果实轮纹病抗病性21.34mm，短枝型0.99；
按照权利要求3～5任意一项所述的方法得到待测苹果属植物样本的基因型；
根据所述待测苹果属植物样本的基因型，按照所述的群体平均值和权利要求1所述分子标记的每个基因型对应性状的基因型效应值或基因型组合效应值，利用如下标准判定待测苹果属植物样本的性状表型或者使用如下预测模型计算待测苹果属植物样本的性状的预测表型值；
权利要求1所述分子标记的每个基因型对应性状的基因型效应值或基因型组合效应值，如表2所示：
表2每个分子标记的每个基因型对相应性状的基因型效应值
所述标准包括：
(1)炭疽叶枯病抗病性：
当S1202的基因型为CC且zhwy64的基因型为CC时，判定为抗病；其它基因型判定为感病；
(2)果形：
当newdy202的基因型为CC、SZ2270的基因型为GG、SZ5253的基因型为CC、SZ9100的基因型为GG或者SZ9195的基因型为AA时，判定果形为圆锥-圆；
当SP031的基因型为CC、SP081的基因型不为CT或者XDY231的基因型为GG时，判定果形为扁圆-圆；
所述预测模型包括：
(3)绿原酸含量或原花青素B2含量采用基因型组合模型；
按照绿原酸的分子标记的基因型或原花青素B2含量的分子标记的基因型组合估算效应值，用基因型组合效应值建立GAP模型，公式为：
GPV＝α×(GcE+μ)+β；
GPV为预测的表型值；GcE为该性状各个标记的基因型组合效应值；μ为训练群体该性状的表型均值；α和β分别为线性回归系数和剩余参数；
(4)果实成熟期、可溶性固形物含量、果汁pH值、采收时果肉硬度、采收时果肉脆度、果肉硬度保持性、果肉脆度保持性或果实轮纹病抗病性采用加性模型，公式为：
GPV为预测的表型值；GE为该性状标记的基因型效应值；k为该性状的标记数；μ为训练群体该性状的表型均值；α和β分别为线性回归系数和剩余参数；
(5)平均单果重、苹果酸含量、果面着色度和短枝型采用固定效应模型，预测公式为：
GPV为预测的表型值；Fx为固定效应标记基因型效应值；GnE为该性状非固定效应标记的基因型效应值；k为该性状的非固定效应的标记数；μ为训练群体该性状的表型均值；γ为缩值系数；α和β分别为线性回归系数和剩余参数；
平均单果重的固定效应为：XDY160的基因型为AA或SZ4849的基因型为GG或SZ4161的基因型为GG，Fx分别为-104.8，-100.7和-101.4；
苹果酸含量的固定效应为Ma、MA202和SAUR-5的基因型组合效应值；
果面着色度的固定效应分别为ZZZ162与zwy6和ZZZ162与color1245的基因型组合效应值；
短枝型的固定效应为neww45与S1245和neww45与ww19的基因型组合效应值；
7.权利要求1所述苹果属植物性状相关分子标记组合或者权利要求2所述引物组合的如下一种或几种应用：
1)苹果属植物性状鉴定；2)苹果属植物指纹图谱或分子身份证构建；3)苹果属种质资源基因型鉴定；4)苹果属植物杂交育种；5)苹果属植物新品种分子DUS测试；
所述苹果属植物杂交育种包括：杂交亲本的选择与杂交组合的选配、杂交世代数设计和杂种后代分子辅助选择中的一种或几种。
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