CN110592104A - 亚麻荠CsDGAT基因及其在植物抗盐方面的应用 - Google Patents

亚麻荠CsDGAT基因及其在植物抗盐方面的应用 Download PDF

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CN110592104A
CN110592104A CN201911027615.2A CN201911027615A CN110592104A CN 110592104 A CN110592104 A CN 110592104A CN 201911027615 A CN201911027615 A CN 201911027615A CN 110592104 A CN110592104 A CN 110592104A
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苑丽霞
李润植
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Abstract

本发明植物抗盐基因技术领域，具体涉及一种亚麻荠CsDGAT基因及其在植物抗盐方面的应用。所述亚麻荠基因CsDGAT为CsDGAT1‑B，CsDGAT1‑B的核苷酸序列如SEQ ID NO.1所示，氨基酸序列如SEQ ID NO.2所示。CsDGAT基因的功能分析和鉴定为获得植物新品种提供了理论基础，利用该基因进行分子育种有望培养出具有生产应用价值的高油植物；为培育抗盐的转基因作物新品种奠定了理论和生产实践基础。

Description

亚麻荠CsDGAT基因及其在植物抗盐方面的应用
技术领域
本发明属于植物抗盐基因技术领域，具体涉及一种亚麻荠CsDGAT基因及其在植物抗盐方面的应用。
背景技术
亚麻荠(Camelina sativa L.Crantz)是十字花科亚麻荠属一年生草本植物。亚麻荠种子产量600～3960kg/hm2，株高30～120cm，平均高度92.4cm，生育期为90天左右。千粒质量平均1.25g，种子含油量高，我国近年来也开展了亚麻荠的引种、育种及推广种植。亚麻荠种子在不同环境中含油量的变化范围是30％～43％，不同基因型间含油量的变化范围是30.8％～32.9％。亚麻荠油中含有高达90％的多种健康有益型的不饱和脂肪酸。亚麻荠种子油突出优点是含有多种对人体有益的独特的不饱和脂肪酸组成，尤其是高水平的α-亚麻酸(ω-3脂肪酸)，是应用于人类营养、心血管及大脑等保健和功能食品生产等的珍贵高值脂肪酸，所以亚麻荠是一种重新发掘的优质能源作物。在不断引进、试种国外品种的同时，持续培育亚麻荠优良品种和建立高效低耗的规模化种植技术体系，全面评估在新种植区域条件下亚麻荠生长发育和种子品质等农艺性状就成为一项迫切任务。
植物的抗盐性是一个受多种因素影响的较为复杂的综合性状，不同植物由于其抗盐机理不同使得其生理代谢和生化变化也不同。高盐是影响植物生长的主要因素，在盐胁迫下，植物体内的主要生理过程都会发生变化。亚麻荠是一种低投入、耐逆境的油料作物，亚麻荠因具有高产、优质、多抗等优良特性，是十字花科作物遗传改良的重要种质资源，更符合可持续农业的发展要求，是一种较为理想的型能源作物。在抗盐品种的选育方面，最常用的方法就是常规杂交育种，常规杂交育种是通过不同基因型个体之间的交配而取得双亲基因重新组合的个体的方法。杂交过程并不会产生新基因，且杂交后代会出现性状分离，育种过程缓慢，过程复杂。因此开发能够鉴定亚麻荠抗盐性的基因，对于亚麻荠抗盐品种的选育具有重要意义。
发明内容
本发明提供了一种亚麻荠CsDGAT基因及其在植物抗盐方面的应用，CsDGAT1-B基因能使亚麻荠的抗盐性能提高。
本发明的目的是提供一种亚麻荠基因CsDGAT，所述亚麻荠基因CsDGAT为CsDGAT1-B，CsDGAT1-B的核苷酸序列如SEQ ID NO.1所示，氨基酸序列如SEQ ID NO.2所示；所述亚麻荠基因CsDGAT能够鉴定植物抗盐性。
本发明还提供了一种所述的亚麻荠基因CsDGAT在植物抗盐育种中的应用。
本发明还提供了一种所述的亚麻荠基因CsDGAT在高油脂含量植物育种中的应用。
优选的，根据上述应用，所述亚麻荠基因CsDGAT还包括CsDGAT1-A、CsDGAT1-C、CsDGAT2-A、CsDGAT2-B和CsDGAT2-C。
优选的，根据上述应用，所述植物为亚麻荠和烟草。
与现有技术相比，本发明提供的一种亚麻荠CsDGAT基因及其在植物抗盐方面的应用，至少具有以下有益效果：
CsDGAT基因的功能分析和鉴定为获得植物新品种提供了理论基础，利用该基因进行分子育种有望培养出具有生产应用价值的高油植物。为培育抗盐的转基因作物新品种奠定了理论和生产实践基础。
我们的研究结果显示盐胁迫处理导致CsDGAT1-B mRNA表达增加15倍，同时也分离了这些CsDGAT基因的cDNA克隆，用于烟草叶片的瞬时表达，分析显示六个基因DGAT酶活性和油脂积累大幅增加。
附图说明
图1是亚麻荠种子发育不同时期CsDGAT基因的表达模式；
图1A对应的是CsDGAT1-A、CsDGAT1-B、CsDGAT1-C基因，图1B对应的是CsDGAT2-A、CsDGAT2-B和CsDGAT2-C；
图2是NaCl胁迫条件下亚麻荠幼苗中TAG的积累；
图3是NaCl胁迫条件下三周龄亚麻荠幼苗CsDGAT基因的表达模式；
图4是CsDGAT基因导入烟草叶片瞬间表达后TAG的含量。
具体实施方式
为了使本领域技术人员更好地理解本发明的技术方案能予以实施，下面结合具体实施例和附图对本发明作进一步说明。
下面各实施例以及上述发明内容中未注明具体条件的试验方法，均按照本领域的常规方法和条件进行，所用的材料若无特殊说明均为市售。
CsDGAT能够编码二酰甘油酰基转移酶(DGAT)，DGAT是参与生成TAG的最后一步酰基化反应的重要酶，该酶表达量越高，则说明亚麻荠的产油脂性能越强。在动植物中，至少发现有两类DGATs,即DGAT1和DGAT2，尽管二者不具序列同源性，但均能催化将结合于CoA的酰基转移到DAG分子的sn-3位置生成TAG酶促反应。
本发明提供了一种能够鉴定植物抗盐性的亚麻荠基因CsDGAT，所述亚麻荠基因CsDGAT为CsDGAT1-B，CsDGAT1-B的核苷酸序列如SEQ ID NO.1所示，氨基酸序列如SEQ IDNO.2所示。CsDGAT1-B在细胞内的第19号染色体上，且有6个外显子，基因的CDS长度为2069bp，编码520个氨基酸的蛋白。
本发明还提供了一种如SEQ ID NO.1所示CsDGAT1-B基因在植物抗盐性育种、尤其是亚麻荠抗盐育种中的应用。
实施例1 CsDGAT基因成员的特征分析
为了鉴定亚麻荠(C.sativa)中编码DGAT的基因，我们使用拟南芥AtDGAT1(AT2G19450)和AtDGAT2(AT3G51520)的氨基酸(AA)序列作为查询序列，对camelina基因组数据库(https://www.ncbi.nlm.nih.gov/genome/？term＝camelina+sativa)进行BLAST搜索。鉴定出6个与拟南芥同源蛋白高度一致的预测蛋白(蛋白同源性>90％)。在亚麻荠基因组中，DGAT1和DGAT2家族各鉴定出3个成员，其中3个成员具有非常高的氨基酸序列相似性(>95％)。CsDGAT1家族包括CsDGAT1-A(XP_010415368，氨基酸序列如SEQ ID NO.3)、CsDGAT1-B(XP_010489480)、CsDGAT1-C(XP_010468809，氨基酸序列如SEQ ID NO.4)三个成员，蛋白大小分别为520、520、474个氨基酸。对于CsDGAT2家族，三个成员分别为CsDGAT2-A(XP_010426724，氨基酸序列如SEQ ID NO.5)、CsDGAT2-B(XP_010515570，氨基酸序列如SEQID NO.6)和CsDGAT2-C(XP_010503848，氨基酸序列如SEQ ID NO.7)，蛋白大小相同，均为315个氨基酸。表1列出了已鉴定的CsDGAT基因的一般信息。通过与已公布的DGATs基因进行综合分析证明，CsDGAT1-A、CsDGAT1-B、CsDGAT1-C、CsDGAT2-A、CsDGAT2-B和CsDGAT2-C具有DGAT家族的明显特征，即均能催化结合CoA的酰基转移到DAG分子的sn-3位置生成TAG的酶促反应。
表1 CsDGAT基因和编码多肽的基本信息
实施例2 CsDGAT1-B基因在盐胁迫中发挥重要作用
采用qRT-PCR检测亚麻荠种子不同发育阶段，即开花后的第7d、15d、22d、29d、36d，CsDGAT1-A、CsDGAT1-B、CsDGAT1-C、CsDGAT2-A、CsDGAT2-B和CsDGAT2-C 6个靶基因的mRNA水平，结果参见图1。CsDGAT1s家族中主要为CsDGAT1-A的mRNA表达量较大，其次为CsDGAT2s家族中的CsDGAT2-C，其中CsDGAT1-A转录本的表达量比CsDGAT2-C高2倍。另一方面，CsDGAT1-A和CsDGAT2-C在开花后的第15～22d种子中表达大量增加，这一时期亚麻荠的油脂积累显著增强，说明CsDGAT1-A和CsDGAT2-C对油脂积累有重要作用。
为了确定CsDGAT1和CsDGAT2是否参与亚麻荠对盐胁迫的响应，收获了在盐胁迫下生长的3周龄亚麻荠幼苗，幼苗样本被分为两批：一份用于总脂质提取和三酰甘油(TAG)测量，另一份用于总RNA提取和qRT-PCR。以盐胁迫处理0h为对照，在150mM浓度盐胁迫处理24h条件下TAG积累逐渐增加到最大水平高于对照1.9倍，结果参见图2，其中从12h到24h发生了TAG快速增长。
同时我们也分析了上述盐胁迫条件下亚麻荠幼苗CsDGAT1-A、CsDGAT1-B、CsDGAT1-C、CsDGAT2-A、CsDGAT2-B和CsDGAT2-C的差异表达，结果参见图3，CsDGAT1-B在12h和24h转录水平大幅度上调至峰值，高于对照15倍，然而CsDGAT1-C和CsDGAT2-A的mRNA表达量在盐胁迫压力中下降，其他基因成员表达没有显著变化。结果分析表明，CsDGAT1-B的表达与盐胁迫条件下亚麻荠种子含油量增加密切相关。
此外，我们还分离了这些CsDGAT1-A、CsDGAT1-B、CsDGAT1-C、CsDGAT2-A、CsDGAT2-B和CsDGAT2-C基因的cDNA克隆，分别将其导入烟草叶片，具体步骤如下：用RT-PCR方法从亚麻荠发育组织的总RNA中扩增上述基因的cDNA，然后分别插入克隆载体。用高保真Taq DNA聚合酶扩增每个ORF序列，表2列出了CsDGAT cDNAs的ORF 5'和3'末端序列的引物。在每个正向和反向引物上分别添加HindIII和BamH1的限制性位点。再将每个CsDGAT基因的相应ORF插入到HindIII-BamH1消化的pBI121载体中，以产生35S为启动子驱动每个CsDGAT基因的表达载体。然后将每个CsDGAT表达载体分别转移到农杆菌GV3101中。农杆菌细胞在28℃下培养过夜。使用渗透缓冲液(10mM MES,pH 5.7,10mM MgCl2,and 150mMacetosyringone)收集并重新悬浮细胞，为使CsDGATs瞬时表达随后用真空渗透法分别对4-5周龄的烟草叶片进行处理，7天后采集烟草叶片(感染的和对应未感染的区域)进行脂质分析。结果参见图4。图4中，WT是野生型烟草叶片中TAG积累量，VC表示pBI121空载体烟草叶片中TAG积累量，CsDGAT1-A、CsDGAT1-B、CsDGAT1-C、CsDGAT2-A、CsDGAT2-B和CsDGAT2-C分别表示烟草叶片中转化了CsDGAT1-A、CsDGAT1-B、CsDGAT1-C、CsDGAT2-A、CsDGAT2-B和CsDGAT2-C基因的TAG积累量。结果表明所有CsDGATs基因的瞬时表达导致烟草叶片总含油量显著增加，且与未感染区(对照WT)和空载体(VC)相比，感染区的TAG水平提高了2倍。
表2用于扩增CsDGATs cDNA的引物序列
需要说明的是本发明权利要求书中涉及数值范围时，应理解为每个数值范围的两个端点以及两个端点之间任何一个数值均可选用，由于采用的步骤方法与实施例相同，为了防止赘述，本发明的描述了优选的实施例，但本领域内的技术人员一旦得知了基本创造性概念，则可对这些实施例作出另外的变更和修改。所以，所附权利要求意欲解释为包括优选实施例以及落入本发明范围的所有变更和修改。显然，本领域的技术人员可以对本发明进行各种改动和变型而不脱离本发明的精神和范围。这样，倘若本发明的这些修改和变型属于本发明权利要求及其等同技术的范围之内，则本发明也意图包含这些改动和变型在内。
序列表
<110> 晋中学院
<120> 亚麻荠CsDGAT基因及其在植物抗盐方面的应用
<160> 21
<170> PatentIn version 3.3
<210> 1
<211> 2069
<212> DNA
<213> 人工序列
<400> 1
taaacaaaca aaccttctct taatatgtac aaacttttat tattctctct ctctattgga 60
ttctgattct cttcaacgtg actcctcacc ttttctttca atttcatctg catgtttctc 120
gattctctct gacgccttcc ttctataacg tttctttgaa agctgtttcc gtcaattcgg 180
taaatggcga ttttggattc tggaggcggc ggcgttagca ccgcgacggc gacagagaac 240
ggtggcggag agtttgtgga tcttcgtcga cggaaatcga gatcggattc caacggagtt 300
ctttctggtt ccgataatcc accgtctgtt gatgttggag ctcccgccga cgttagggat 360
cggattgatt ccgttgttaa cgatgacgct caggggacga ctgccaattt ggccggagat 420
accgaaatta gggaaaccgg tggtggtgga agaggcgccg gcggagaagg aggaagaggt 480
aacgccgaga ctacgtatgc gtatcgaccg tcggttcctg ctcatcggag agctagggag 540
agtccactca gctccgacgc aatcttcaaa cagagccatg ccggattatt caacttgtgc 600
gtagtagttc ttattgctgt aaacagtaga ctcatcatcg aaaatctgat gaagtatggt 660
tggttgatca gaacggattt ctggtttagt tcaagatcgt tgcgggattg gccgcttttc 720
atgtgttgtc tctccctttc aatctttcct ttggctgcct ttaccgtcga gaaattggtt 780
cttcagaaat gcatttctga acctgttgtc atcattcttc atattattat caccatgaca 840
gaggttttgt atccagttta cgtcacccta aggtgtgatt ctgccttctt atcaggtgtc 900
acattgatgc tcctcacttg cattgtgtgg ctaaagttgg tttcttatgc tcatactaac 960
tacgacataa gaactctagc caattcagct gataaggcca atcctgaagt ctcctactac 1020
gttagcttga agagcttggc atattttatg gttgctccca cattgtgtta tcagccaagc 1080
tatccacgtt ctccatgtat acggaagggt tgggtggctc gtcaatttgc aaaactggtc 1140
atattcactg gattcatggg atttataata gaacaatata taaatcctat tgtcagaaac 1200
tcaaagcatc ctctgaaagg ggatcttcta tatgctattg aaagagtgtt gaagctttca 1260
gttccaaatt tatatgtgtg gctctgcatg ttctactgct tcttccacct ttggttaaac 1320
atattggcag agcttctctg cttcggggat cgtgaattct acagagattg gtggaatgca 1380
aaaagtgtgg gagactattg gagaatgtgg aatatgcctg ttcataaatg gatggttcga 1440
catatatact tcccgtgcct gcggagcaag ataccaaaga cactcgccat tatcattgct 1500
ttcttagtct ctgccgtctt tcatgagcta tgcatcgcag tcccttgccg tctcttcaag 1560
ttatgggctt ttatagggat tatgtttcag gtgcctttgg tctttatcac aaactatcta 1620
caagaaaggt tcggctcaac ggtggggaac atgatcttct ggttcatctt ctgcatattc 1680
ggacaaccga tgtgtgtgct tctttattac cacgatctga tgaaccgcaa aggatcaatg 1740
tcatgaaaca actgttggaa attgactttc tccaaacata tgtggttgtg atgctaaaac 1800
aagaatagtg ttttaaccat tgaagaagaa aaaaaaaagt taaaaaatta gagttgttgt 1860
atctgcaaaa ttttggtaga gacgaaccgt ttttgagttt tgaatctcgt tgtcaagtct 1920
ctcttttgat cttgctatgc tcttgtcttt aaaccacaat caagcaaagt agattgagat 1980
attgaatttt gttttggtgt aaagaaaatt tcaatcaaaa actgttgtaa taattggtgc 2040
aaaaaaaatt aataaatggt ttttttttt 2069
<210> 2
<211> 520
<212> PRT
<213> 人工序列
<400> 2
Met Ala Ile Leu Asp Ser Gly Gly Gly Gly Val Ser Thr Ala Thr Ala
1 5 10 15
Thr Glu Asn Gly Gly Gly Glu Phe Val Asp Leu Arg Arg Arg Lys Ser
20 25 30
Arg Ser Asp Ser Asn Gly Val Leu Ser Gly Ser Asp Asn Pro Pro Ser
35 40 45
Val Asp Val Gly Ala Pro Ala Asp Val Arg Asp Arg Ile Asp Ser Val
50 55 60
Val Asn Asp Asp Ala Gln Gly Thr Thr Ala Asn Leu Ala Gly Asp Thr
65 70 75 80
Glu Ile Arg Glu Thr Gly Gly Gly Gly Arg Gly Ala Gly Gly Glu Gly
85 90 95
Gly Arg Gly Asn Ala Glu Thr Thr Tyr Ala Tyr Arg Pro Ser Val Pro
100 105 110
Ala His Arg Arg Ala Arg Glu Ser Pro Leu Ser Ser Asp Ala Ile Phe
115 120 125
Lys Gln Ser His Ala Gly Leu Phe Asn Leu Cys Val Val Val Leu Ile
130 135 140
Ala Val Asn Ser Arg Leu Ile Ile Glu Asn Leu Met Lys Tyr Gly Trp
145 150 155 160
Leu Ile Arg Thr Asp Phe Trp Phe Ser Ser Arg Ser Leu Arg Asp Trp
165 170 175
Pro Leu Phe Met Cys Cys Leu Ser Leu Ser Ile Phe Pro Leu Ala Ala
180 185 190
Phe Thr Val Glu Lys Leu Val Leu Gln Lys Cys Ile Ser Glu Pro Val
195 200 205
Val Ile Ile Leu His Ile Ile Ile Thr Met Thr Glu Val Leu Tyr Pro
210 215 220
Val Tyr Val Thr Leu Arg Cys Asp Ser Ala Phe Leu Ser Gly Val Thr
225 230 235 240
Leu Met Leu Leu Thr Cys Ile Val Trp Leu Lys Leu Val Ser Tyr Ala
245 250 255
His Thr Asn Tyr Asp Ile Arg Thr Leu Ala Asn Ser Ala Asp Lys Ala
260 265 270
Asn Pro Glu Val Ser Tyr Tyr Val Ser Leu Lys Ser Leu Ala Tyr Phe
275 280 285
Met Val Ala Pro Thr Leu Cys Tyr Gln Pro Ser Tyr Pro Arg Ser Pro
290 295 300
Cys Ile Arg Lys Gly Trp Val Ala Arg Gln Phe Ala Lys Leu Val Ile
305 310 315 320
Phe Thr Gly Phe Met Gly Phe Ile Ile Glu Gln Tyr Ile Asn Pro Ile
325 330 335
Val Arg Asn Ser Lys His Pro Leu Lys Gly Asp Leu Leu Tyr Ala Ile
340 345 350
Glu Arg Val Leu Lys Leu Ser Val Pro Asn Leu Tyr Val Trp Leu Cys
355 360 365
Met Phe Tyr Cys Phe Phe His Leu Trp Leu Asn Ile Leu Ala Glu Leu
370 375 380
Leu Cys Phe Gly Asp Arg Glu Phe Tyr Arg Asp Trp Trp Asn Ala Lys
385 390 395 400
Ser Val Gly Asp Tyr Trp Arg Met Trp Asn Met Pro Val His Lys Trp
405 410 415
Met Val Arg His Ile Tyr Phe Pro Cys Leu Arg Ser Lys Ile Pro Lys
420 425 430
Thr Leu Ala Ile Ile Ile Ala Phe Leu Val Ser Ala Val Phe His Glu
435 440 445
Leu Cys Ile Ala Val Pro Cys Arg Leu Phe Lys Leu Trp Ala Phe Ile
450 455 460
Gly Ile Met Phe Gln Val Pro Leu Val Phe Ile Thr Asn Tyr Leu Gln
465 470 475 480
Glu Arg Phe Gly Ser Thr Val Gly Asn Met Ile Phe Trp Phe Ile Phe
485 490 495
Cys Ile Phe Gly Gln Pro Met Cys Val Leu Leu Tyr Tyr His Asp Leu
500 505 510
Met Asn Arg Lys Gly Ser Met Ser
515 520
<210> 3
<211> 520
<212> PRT
<213> 人工序列
<400> 3
Met Ala Ile Leu Asp Ser Gly Gly Gly Gly Val Ser Thr Ala Thr Ala
1 5 10 15
Thr Glu Asn Gly Gly Gly Glu Phe Val Asp Leu Arg Arg Arg Lys Ser
20 25 30
Arg Ser Asp Ser Asn Gly Val Leu Ser Gly Ser Asp Asn Pro Pro Ser
35 40 45
Val Asp Val Gly Ala Pro Ala Asp Val Arg Asp Arg Ile Asp Ser Val
50 55 60
Val Asn Asp Asp Ala Gln Gly Thr Thr Ala Asn Leu Ala Gly Asp Thr
65 70 75 80
Glu Ile Arg Glu Thr Gly Gly Gly Gly Arg Gly Ala Gly Gly Glu Gly
85 90 95
Gly Arg Gly Asn Ala Glu Thr Thr Tyr Ala Tyr Arg Pro Ser Val Pro
100 105 110
Ala His Arg Arg Ala Arg Glu Ser Pro Leu Ser Ser Asp Ala Ile Phe
115 120 125
Lys Gln Ser His Ala Gly Leu Phe Asn Leu Cys Val Val Val Leu Ile
130 135 140
Ala Val Asn Ser Arg Leu Ile Ile Glu Asn Leu Met Lys Tyr Gly Trp
145 150 155 160
Leu Ile Arg Thr Asp Phe Trp Phe Ser Ser Arg Ser Leu Arg Asp Trp
165 170 175
Pro Leu Phe Met Cys Cys Leu Ser Leu Ser Ile Phe Pro Leu Ala Ala
180 185 190
Phe Thr Val Glu Lys Leu Val Leu Gln Lys Cys Ile Ser Glu Pro Val
195 200 205
Val Ile Ile Leu His Ile Ile Ile Thr Met Thr Glu Val Leu Tyr Pro
210 215 220
Val Tyr Val Thr Leu Arg Cys Asp Ser Ala Phe Leu Ser Gly Val Thr
225 230 235 240
Leu Met Leu Leu Thr Cys Ile Val Trp Leu Lys Leu Val Ser Tyr Ala
245 250 255
His Thr Asn Tyr Asp Ile Arg Thr Leu Ala Asn Ser Ala Asp Lys Ala
260 265 270
Asn Pro Glu Val Ser Tyr Tyr Val Ser Leu Lys Ser Leu Ala Tyr Phe
275 280 285
Met Val Ala Pro Thr Leu Cys Tyr Gln Pro Ser Tyr Pro Arg Ser Pro
290 295 300
Cys Ile Arg Lys Gly Trp Val Ala Arg Gln Phe Ala Lys Leu Val Ile
305 310 315 320
Phe Thr Gly Phe Met Gly Phe Ile Ile Glu Gln Tyr Ile Asn Pro Ile
325 330 335
Val Arg Asn Ser Lys His Pro Leu Lys Gly Asp Leu Leu Tyr Ala Ile
340 345 350
Glu Arg Val Leu Lys Leu Ser Val Pro Asn Leu Tyr Val Trp Leu Cys
355 360 365
Met Phe Tyr Cys Phe Phe His Leu Trp Leu Asn Ile Leu Ala Glu Leu
370 375 380
Leu Cys Phe Gly Asp Arg Glu Phe Tyr Arg Asp Trp Trp Asn Ala Lys
385 390 395 400
Ser Val Gly Asp Tyr Trp Arg Met Trp Asn Met Pro Val His Lys Trp
405 410 415
Met Val Arg His Ile Tyr Phe Pro Cys Leu Arg Ser Lys Ile Pro Lys
420 425 430
Thr Leu Ala Ile Ile Ile Ala Phe Leu Val Ser Ala Val Phe His Glu
435 440 445
Leu Cys Ile Ala Val Pro Cys Arg Leu Phe Lys Leu Trp Ala Phe Ile
450 455 460
Gly Ile Met Phe Gln Val Pro Leu Val Phe Ile Thr Asn Tyr Leu Gln
465 470 475 480
Glu Arg Phe Gly Ser Thr Val Gly Asn Met Ile Phe Trp Phe Ile Phe
485 490 495
Cys Ile Phe Gly Gln Pro Met Cys Val Leu Leu Tyr Tyr His Asp Leu
500 505 510
Met Asn Arg Lys Gly Ser Met Ser
515 520
<210> 4
<211> 474
<212> PRT
<213> 人工序列
<400> 4
Met Ala Ile Leu Asp Ser Gly Gly Gly Gly Val Ser Thr Ala Thr Ala
1 5 10 15
Thr Glu Asn Gly Gly Gly Glu Phe Val Asp Leu Arg Arg Arg Lys Thr
20 25 30
Arg Ser Asp Ser Asn Gly Val Leu Ser Gly Ser Asp Asn Pro Pro Ser
35 40 45
Asp Asp Val Gly Ala Pro Ala Asp Val Arg Asp Arg Ile Asp Ser Val
50 55 60
Val Asn Asp Asp Ala Gln Gly Thr Thr Ala Asn Leu Ala Gly Asp Asn
65 70 75 80
Glu Ile Arg Glu Thr Gly Gly Gly Gly Gly Arg Gly Gly Gly Gly Glu
85 90 95
Gly Gly Arg Gly Asn Ala Glu Thr Thr Tyr Thr Tyr Arg Pro Ser Val
100 105 110
Pro Ala His Arg Arg Ala Arg Glu Ser Pro Leu Ser Ser Asp Ala Ile
115 120 125
Phe Lys Gln Ser His Ala Gly Leu Phe Asn Leu Cys Val Val Val Leu
130 135 140
Ile Ala Val Asn Ser Arg Leu Ile Ile Glu Asn Leu Met Lys Tyr Gly
145 150 155 160
Trp Leu Ile Arg Thr Asp Phe Trp Phe Ser Ser Arg Ser Leu Arg Asp
165 170 175
Trp Pro Leu Phe Met Cys Cys Leu Ser Leu Ser Phe Phe Pro Leu Ala
180 185 190
Ala Phe Thr Val Glu Lys Leu Val Leu Gln Lys Cys Ile Ser Glu Pro
195 200 205
Val Val Ile Phe Leu His Ile Ile Ile Thr Met Thr Glu Val Leu Tyr
210 215 220
Pro Val Tyr Val Thr Leu Arg Cys Asp Ser Ala Phe Leu Ser Gly Val
225 230 235 240
Thr Leu Met Leu Leu Thr Cys Ile Val Trp Leu Lys Leu Val Ser Tyr
245 250 255
Ala His Thr Asn Tyr Asp Ile Arg Thr Leu Ala Asn Ser Ala Asp Lys
260 265 270
Val Glu Glu Glu Asn Phe Met Gly Phe Ile Ile Glu Gln Tyr Ile Asn
275 280 285
Pro Ile Val Arg Asn Ser Lys His Pro Leu Lys Gly Asp Leu Leu Tyr
290 295 300
Ala Ile Glu Arg Val Leu Lys Leu Ser Val Pro Asn Leu Tyr Val Trp
305 310 315 320
Leu Cys Met Phe Tyr Cys Phe Phe His Leu Trp Leu Asn Ile Leu Ala
325 330 335
Glu Leu Leu Cys Phe Gly Asp Arg Glu Phe Tyr Arg Asp Trp Trp Asn
340 345 350
Ala Lys Ser Val Gly Asp Tyr Trp Arg Met Trp Asn Met Pro Val His
355 360 365
Lys Trp Met Val Arg His Ile Tyr Phe Pro Cys Leu Arg Ser Lys Ile
370 375 380
Pro Lys Thr Leu Ala Ile Ile Ile Ala Phe Leu Val Ser Ala Val Phe
385 390 395 400
His Glu Leu Cys Ile Ala Val Pro Cys Arg Leu Phe Lys Leu Trp Ala
405 410 415
Phe Ile Gly Ile Met Phe Gln Val Pro Leu Val Phe Ile Thr Asn Tyr
420 425 430
Leu Gln Glu Arg Phe Gly Ser Thr Val Gly Asn Met Ile Phe Trp Phe
435 440 445
Ile Phe Cys Ile Phe Gly Gln Pro Met Cys Val Leu Leu Tyr Tyr His
450 455 460
Asp Leu Met Asn Arg Lys Gly Ser Met Ala
465 470
<210> 5
<211> 315
<212> PRT
<213> 人工序列
<400> 5
Met Gly Gly Ser Arg Glu Phe Arg Ala Asp Lys Ser Ser Asp Gln Phe
1 5 10 15
His Ser Thr Ile Ala Met Ala Ile Trp Leu Gly Ala Ile His Phe Asn
20 25 30
Ile Val Leu Val Leu Phe Ser Leu Ile Phe Leu Pro Pro Tyr Leu Ser
35 40 45
Leu Leu Val Leu Ser Leu Leu Ser Leu Phe Ile Phe Ile Pro Ile Asp
50 55 60
His Arg Ser Lys Tyr Gly Arg Lys Leu Ala Arg Tyr Ile Cys Lys His
65 70 75 80
Ala Cys Asn Tyr Phe Pro Val Ser Leu Tyr Val Glu Asp Tyr Glu Ala
85 90 95
Phe Gln Pro Thr Arg Ala Tyr Val Phe Gly Tyr Glu Pro His Ser Val
100 105 110
Leu Pro Ile Gly Val Val Ala Leu Cys Asp Leu Thr Gly Phe Met Pro
115 120 125
Leu Thr Lys Ile Lys Val Leu Ala Ser Ser Ala Ile Phe Tyr Thr Pro
130 135 140
Phe Leu Arg His Ile Trp Thr Trp Leu Gly Leu Thr Ala Ala Ser Arg
145 150 155 160
Lys Asn Phe Thr Ser Leu Leu Asp Ser Gly Tyr Ser Cys Val Leu Val
165 170 175
Pro Gly Gly Val Gln Glu Thr Phe His Met Gln His Asp Val Glu Asn
180 185 190
Val Phe Leu Ser Ser Arg Arg Gly Phe Val Arg Ile Ala Met Glu Gln
195 200 205
Gly Thr Pro Ile Val Pro Val Phe Cys Phe Gly Gln Ser His Val Tyr
210 215 220
Lys Trp Trp Lys Pro Asp Trp Asp Leu Tyr Leu Lys Leu Ser Arg Ala
225 230 235 240
Ile Arg Phe Thr Pro Ile Cys Phe Gly Gly Val Phe Gly Ser Pro Leu
245 250 255
Pro Tyr Arg Gln Pro Met His Val Ile Val Gly Lys Pro Ile Glu Val
260 265 270
Thr Lys Thr Leu Gln Pro Thr Asp Glu Glu Ile Ala Lys Phe His Gly
275 280 285
Gln Tyr Val Glu Ala Leu Arg Asp Leu Phe Glu Gly His Lys Ala Arg
290 295 300
Val Gly Tyr Pro Asp Leu Gln Leu Asn Ile Leu
305 310 315
<210> 6
<211> 315
<212> PRT
<213> 人工序列
<400> 6
Met Gly Gly Ser Arg Glu Phe Arg Ala Asp Lys Ser Ser Asp Gln Phe
1 5 10 15
His Ser Thr Ile Ala Met Ala Ile Trp Leu Gly Ala Ile His Phe Asn
20 25 30
Ile Val Leu Val Leu Phe Ser Leu Ile Phe Leu Pro Pro Tyr Leu Ser
35 40 45
Leu Leu Val Leu Ser Leu Leu Ser Leu Phe Ile Phe Ile Pro Ile Asp
50 55 60
His Arg Ser Lys Tyr Gly Arg Lys Leu Ala Arg Tyr Ile Cys Lys His
65 70 75 80
Ala Cys Asn Tyr Phe Pro Val Ser Met Tyr Val Glu Asp Tyr Glu Ala
85 90 95
Phe Gln Pro Asn Arg Ala Tyr Val Phe Gly Tyr Glu Pro His Ser Val
100 105 110
Leu Pro Ile Gly Val Val Ala Leu Cys Asp Leu Thr Gly Phe Met Pro
115 120 125
Leu Thr Lys Ile Lys Val Leu Ala Ser Ser Ala Ile Phe Tyr Thr Pro
130 135 140
Phe Leu Arg His Ile Trp Thr Trp Leu Gly Leu Thr Pro Ala Ser Arg
145 150 155 160
Lys Asn Phe Thr Ser Leu Leu Asp Ser Gly Tyr Ser Cys Val Leu Val
165 170 175
Pro Gly Gly Val Gln Glu Thr Phe His Met Gln His Asp Val Glu Asn
180 185 190
Val Phe Leu Ser Ser Arg Arg Gly Phe Val Arg Ile Ala Met Glu Gln
195 200 205
Gly Thr Pro Leu Val Pro Val Phe Cys Phe Gly Gln Ser His Val Tyr
210 215 220
Arg Trp Trp Lys Pro Asp Trp Asp Leu Tyr Leu Lys Leu Ser Arg Ala
225 230 235 240
Ile Arg Phe Thr Pro Ile Cys Phe Gly Gly Val Phe Gly Ser Pro Leu
245 250 255
Pro Tyr Arg Gln Pro Met His Val Ile Val Gly Lys Pro Ile Glu Val
260 265 270
Thr Lys Thr Leu Gln Pro Thr Asp Glu Glu Ile Ala Lys Phe His Gly
275 280 285
Gln Tyr Val Glu Ala Leu Arg Asp Leu Phe Glu Ser His Lys Ala Arg
290 295 300
Val Gly Tyr Pro Asp Leu Gln Leu Asn Ile Leu
305 310 315
<210> 7
<211> 315
<212> PRT
<213> 人工序列
<400> 7
Met Gly Gly Ser Arg Glu Phe Arg Ala Asp Lys Ser Ser Asp Gln Phe
1 5 10 15
His Ser Thr Ile Ala Met Ala Ile Trp Leu Gly Ala Ile His Phe Asn
20 25 30
Ile Val Leu Val Leu Phe Ser Leu Ile Phe Leu Pro Pro Tyr Leu Ser
35 40 45
Leu Leu Val Leu Ser Leu Leu Ser Leu Phe Ile Phe Ile Pro Ile Asp
50 55 60
His Cys Ser Lys Tyr Gly Arg Lys Leu Ala Arg Tyr Ile Cys Lys His
65 70 75 80
Ala Cys Asn Tyr Phe Pro Val Ser Leu Tyr Val Glu Asp Tyr Glu Ala
85 90 95
Phe Gln Pro Asn Arg Ala Tyr Val Phe Gly Tyr Glu Pro His Ser Val
100 105 110
Leu Pro Ile Gly Val Val Ala Leu Cys Asp Leu Thr Gly Phe Met Pro
115 120 125
Leu Thr Lys Ile Lys Val Leu Ala Ser Ser Ala Ile Phe Tyr Thr Pro
130 135 140
Phe Leu Arg His Ile Trp Thr Trp Leu Gly Leu Thr Pro Ala Ser Arg
145 150 155 160
Lys Asn Phe Thr Ser Leu Leu Asp Ser Gly Tyr Ser Cys Val Leu Val
165 170 175
Pro Gly Gly Val Gln Glu Thr Phe His Met Gln His Asp Val Glu Asn
180 185 190
Val Phe Leu Ser Ser Arg Arg Gly Phe Val Arg Ile Ala Met Glu Gln
195 200 205
Gly Thr Pro Ile Val Pro Val Phe Cys Phe Gly Gln Ser His Val Tyr
210 215 220
Lys Trp Trp Lys Pro Asp Trp Asp Leu Tyr Leu Lys Leu Ser Arg Ala
225 230 235 240
Ile Arg Phe Thr Pro Ile Cys Phe Gly Gly Val Phe Gly Ser Pro Leu
245 250 255
Pro Tyr Arg Gln Pro Met His Val Ile Val Gly Lys Pro Ile Glu Val
260 265 270
Thr Lys Thr Leu Gln Pro Thr Asp Glu Glu Ile Ala Lys Phe His Gly
275 280 285
Gln Tyr Val Glu Ala Leu Arg Asp Leu Phe Glu Ser His Lys Ala Arg
290 295 300
Val Gly Tyr Pro Asp Leu Gln Leu Asn Ile Leu
305 310 315
<210> 8
<211> 19
<212> DNA
<213> 人工序列
<400> 8
caccctctga tgatgttgg 19
<210> 9
<211> 21
<212> DNA
<213> 人工序列
<400> 9
atggctctgt ttgaagattg c 21
<210> 10
<211> 20
<212> DNA
<213> 人工序列
<400> 10
cgagatcgga ttccaacgga 20
<210> 11
<211> 20
<212> DNA
<213> 人工序列
<400> 11
gcatacgtag tctcggcgtt 20
<210> 12
<211> 20
<212> DNA
<213> 人工序列
<400> 12
ttttggattc tggaggcggc 20
<210> 13
<211> 20
<212> DNA
<213> 人工序列
<400> 13
tttcgttatc cccggccaaa 20
<210> 14
<211> 18
<212> DNA
<213> 人工序列
<400> 14
cgccattcac ttcaacat 18
<210> 15
<211> 18
<212> DNA
<213> 人工序列
<400> 15
ccaaagacat aggcacga 18
<210> 16
<211> 18
<212> DNA
<213> 人工序列
<400> 16
aagcggacaa accaaaga 18
<210> 17
<211> 20
<212> DNA
<213> 人工序列
<400> 17
tcgggatgaa gataaacaga 20
<210> 18
<211> 18
<212> DNA
<213> 人工序列
<400> 18
cgccattcac ttcaacat 18
<210> 19
<211> 18
<212> DNA
<213> 人工序列
<400> 19
aaagacatag gcacgatt 18
<210> 20
<211> 24
<212> DNA
<213> 人工序列
<400> 20
acaatttccc gctctgctgt tgtg 24
<210> 21
<211> 24
<212> DNA
<213> 人工序列
<400> 21
agggtttctc tcttccacat gcca 24

Claims (5)

1.一种亚麻荠基因CsDGAT，其特征在于，所述亚麻荠基因CsDGAT为CsDGAT1-B，CsDGAT1-B的核苷酸序列如SEQ ID NO.1所示，氨基酸序列如SEQ ID NO.2所示；所述亚麻荠基因CsDGAT能够鉴定植物抗盐性。
2.根据权利要求1所述的亚麻荠基因CsDGAT在植物抗盐育种中的应用。
3.根据权利要求1所述的亚麻荠基因CsDGAT在高油脂含量植物育种中的应用。
4.根据权利要求3所述的亚麻荠基因CsDGAT在高油脂含量植物育种中的应用，其特征在于，所述亚麻荠基因CsDGAT还包括CsDGAT1-A、CsDGAT1-C、CsDGAT2-A、CsDGAT2-B和CsDGAT2-C。
5.根据权利要求3或4所述的应用，其特征在于，所述植物为亚麻荠和烟草。
CN201911027615.2A 2019-10-28 2019-10-28 亚麻荠CsDGAT基因及其在植物抗盐方面的应用 Pending CN110592104A (zh)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102656267A (zh) * 2009-09-15 2012-09-05 蓝宝石能源公司 耐盐生物
EP2358864B1 (en) * 2008-10-23 2017-08-02 Matrix Genetics, LLC Modified photosynthetic microorganisms for producing triglycerides
CN107129529A (zh) * 2016-02-29 2017-09-05 中国科学院遗传与发育生物学研究所 大豆转录因子GmAREB3在植物油脂代谢调控中的应用
EP2635105B1 (en) * 2010-11-04 2017-12-20 Ben-Gurion University of The Negev Research and Development Authority Acyl-coa: diacylglycerol acyltransferase 1-like gene (ptdgat1) and uses thereof

Patent Citations (4)

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Publication number Priority date Publication date Assignee Title
EP2358864B1 (en) * 2008-10-23 2017-08-02 Matrix Genetics, LLC Modified photosynthetic microorganisms for producing triglycerides
CN102656267A (zh) * 2009-09-15 2012-09-05 蓝宝石能源公司 耐盐生物
EP2635105B1 (en) * 2010-11-04 2017-12-20 Ben-Gurion University of The Negev Research and Development Authority Acyl-coa: diacylglycerol acyltransferase 1-like gene (ptdgat1) and uses thereof
CN107129529A (zh) * 2016-02-29 2017-09-05 中国科学院遗传与发育生物学研究所 大豆转录因子GmAREB3在植物油脂代谢调控中的应用

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NCBI: "XM_010491178.1", 《GENBANK》 *
YUAN LIXIA等: "Spatio-temporal expression and stress responses of DGAT1, DGAT2 and PDAT responsible for TAG biosynthesis in Camelina sativa", 《EMIRATES JOURNAL OF FOOD AND AGRICULTURE》 *
郝敬云等: "莱茵衣藻DGAT2基因家族的鉴定与功能分析 ", 《分子植物育种》 *

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Application publication date: 20191220