CN108299549B - Bs1蛋白在调控植物细胞壁木聚糖乙酰化修饰水平中的应用 - Google Patents
Bs1蛋白在调控植物细胞壁木聚糖乙酰化修饰水平中的应用 Download PDFInfo
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- CN108299549B CN108299549B CN201710020763.6A CN201710020763A CN108299549B CN 108299549 B CN108299549 B CN 108299549B CN 201710020763 A CN201710020763 A CN 201710020763A CN 108299549 B CN108299549 B CN 108299549B
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Abstract
本发明提供了BS1蛋白在调控植物细胞壁木聚糖乙酰化修饰水平中的应用。通过实验证明：BS1蛋白具有提高植物细胞壁木聚糖去乙酰化修饰能力的功能，可用于调控植物细胞壁木聚糖的乙酰化修饰水平。对于培育木聚糖乙酰化修饰水平改变的转基因植物具有重大价值，将在生物质资源转变为能源的过程中发挥重大作用，有助于降低生物能源的生产成本，具有重要经济价值。
Description
技术领域
本发明属于生物技术领域，具体涉及BS1蛋白在调控植物细胞壁木聚糖乙酰化修饰水平中的应用。
背景技术
O-乙酰化修饰是植物细胞壁多糖中一种较为普遍的修饰形式。除了纤维素、β-1,3-1,4-连接的葡聚糖和一些糖蛋白，O-乙酰化修饰几乎存在于所有其它细胞壁多糖上。
O-乙酰化修饰可能通过影响细胞壁多糖的生理生化特性从而对细胞壁结构产生影响。研究发现，乙酰化影响多糖与极性分子的相互作用。木聚糖上乙酰化修饰的模式影响了木聚糖与纤维素的结合方式，进而影响细胞壁结构。植物细胞壁多糖O-乙酰化修饰程度随植物组织器官和生长发育阶段呈现动态变化，表明O-乙酰化修饰与植物生长发育状态密切相关，并在植物体内受到严格调控。因此植物细胞壁多糖O-乙酰化修饰的调控机制对于植物维持细胞壁结构与正常生长有重要意义。
研究已发现植物中存在乙酰基转移酶，参与植物细胞壁多糖合成过程中的乙酰化修饰，但该过程中是否存在植物去乙酰化修饰的机制尚未见报道。解析细胞壁多糖的去乙酰化修饰机理是对植物细胞壁结构和生长发育表型进行遗传改良的基础。
在将生物质资源转变为生物能源乙醇的过程中，需要进行微生物发酵，而植物细胞壁多糖释放的乙酰基导致环境酸化，影响发酵过程，而优化条件又增加了乙醇生产的成本。因此细胞壁多糖去乙酰化调控有助于降低生物能源的生产成本，具有重要经济价值。
发明内容
本发明的一个目的是提供如下a)或b)或c)的蛋白质的新用途：
a)氨基酸序列是序列表中序列1所示的蛋白质；
b)在序列表中序列1所示的蛋白质的N端和/或C端连接标签得到的融合蛋白质；
c)将序列表中序列1所示的氨基酸序列经过一个或几个氨基酸残基的取代和/或缺失和/或添加得到的具有相同功能的蛋白质。
本发明提供了上述蛋白质在调控植物细胞壁木聚糖乙酰化修饰水平中的应用。
本发明还提供了上述蛋白质在制备调控植物细胞壁木聚糖乙酰化修饰水平的产品中的应用。
本发明的另一个目的是提供与上述蛋白质相关的生物材料的新用途。
本发明提供了与上述蛋白质相关的生物材料在调控植物细胞壁木聚糖乙酰化修饰水平中的应用。
本发明还提供了与上述蛋白质相关的生物材料在制备调控植物细胞壁木聚糖乙酰化修饰水平的产品中的应用。
所述与上述蛋白质相关的生物材料为下述A1)至A12)中的任一种：
A1)编码上述蛋白质的核酸分子；
A2)含有A1)所述核酸分子的表达盒；
A3)含有A1)所述核酸分子的重组载体；
A4)含有A2)所述表达盒的重组载体；
A5)含有A1)所述核酸分子的重组微生物；
A6)含有A2)所述表达盒的重组微生物；
A7)含有A3)所述重组载体的重组微生物；
A8)含有A4)所述重组载体的重组微生物；
A9)含有A1)所述核酸分子的转基因植物细胞系；
A10)含有A2)所述表达盒的转基因植物细胞系；
A11)含有A3)所述重组载体的转基因植物细胞系；
A12)含有A4)所述重组载体的转基因植物细胞系。
上述应用中，A1)所述核酸分子为如下1)或2)或3)所示的基因：
1)其编码序列是序列表中序列2的cDNA分子或DNA分子；
2)与1)限定的核苷酸序列具有75％或75％以上同一性，且编码上述蛋白质的cDNA分子或基因组DNA分子；
3)在严格条件下与1)或2)限定的核苷酸序列杂交，且编码上述蛋白质的cDNA分子或基因组DNA分子。
上述应用中，所述调控植物细胞壁木聚糖乙酰化修饰水平为提高植物细胞壁木聚糖去乙酰化修饰的能力。
上述应用中，所述提高植物细胞壁木聚糖去乙酰化修饰的能力体现在降低植物细胞壁木聚糖组分中乙酰基含量，具体体现在降低植物细胞壁木聚糖组分中释放的乙酸量。
上述应用中，所述木聚糖乙酰化为木聚糖O-2位和/或O-3位的乙酰化。
本发明还有一个目的是提供上述蛋白质或上述相关生物材料的新用途。
本发明提供了上述蛋白质或上述相关生物材料在培育细胞壁木聚糖乙酰化修饰水平改变的转基因植物中的应用。
上述应用中，所述改变为降低或提高。在本发明的一个实施例中，所述改变具体为降低。
本发明的最后一个目的是提供一种培育细胞壁木聚糖乙酰化修饰水平降低的转基因植物的方法。
本发明提供的培育细胞壁木聚糖乙酰化修饰水平降低的转基因植物的方法包括在受体植物中过表达上述蛋白质，得到转基因植物的步骤；
所述转基因植物细胞壁木聚糖乙酰化修饰水平低于所述受体植物。
上述方法中，所述过表达的方法为将权利要求1中所述的蛋白质的编码基因导入受体植物；
所述转基因植物细胞壁的木聚糖乙酰化修饰水平低于所述受体植物体现在转基因植物细胞壁去乙酰化修饰的能力高于受体植物。
上述方法中，所述蛋白质的编码基因的核苷酸序列是序列2所示的DNA分子。
上述方法中，所述受体植物为单子叶植物或双子叶植物；所述受体植物具体为单子叶植物；所述单子叶植物可为水稻(如粳稻品种“黄金晴”)、玉米(如玉米品种“齐319”)或杨树(如杨树品种“河北杨”)；所述单子叶植物具体为水稻粳稻品种“黄金晴”。
上述方法中，所述木聚糖为乙酰木聚糖。
本发明提供了BS1蛋白在调控植物细胞壁木聚糖去乙酰化修饰中的应用。本发明在野生型水稻黄金晴中过表达BS1蛋白，获得细胞壁木聚糖乙酰化修饰水平降低的转基因植物，而突变体bs1的细胞壁乙酰化修饰水平提高。通过实验证明：BS1蛋白具有提高植物细胞壁木聚糖去乙酰化修饰能力的功能，可用于调控植物细胞壁木聚糖的乙酰化修饰水平。对于培育木聚糖乙酰化修饰水平改变的转基因植物具有重大价值，将在生物质资源转变为能源的过程中发挥重大作用，有助于降低生物能源的生产成本，具有重要经济价值。
附图说明
图1为BS1蛋白的结构示意图。
图2为BS1蛋白对植物木聚糖乙酰化程度的影响。纵坐标为每mg植物茎杆醇不溶物AIR释放的乙酸量(μg)，横坐标依次为总细胞壁(Total)、抽提的果胶组分(Pectin)和其余组分(Remains，木聚糖组分)。其中，NP为野生型植株(粳稻品种“黄金晴”)，bs1为突变体bs1，BS1OE为株系BS1OE的T1代植株。
图3为NMR检测BS1蛋白对木聚糖乙酰化位点的影响。
具体实施方式
下述实施例中所使用的实验方法如无特殊说明，均为常规方法。
下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
下述实施例中的定量试验，均设置三次重复实验，结果取平均值。
下述实施例中的粳稻品种“黄金晴”(WT，又称野生型植株)购买于中国水稻所。
下述实施例中的载体pCAMBIA1300购自CAMIA公司，澳大利亚。
下述实施例中的农杆菌EHA105购自CAMIA公司，澳大利亚。
下述实施例中的载体pDONR207购自Life Technologies公司，货号12213-013。
下述实施例中的去乙酰化修饰是指通过乙酰酯酶特异作用于乙酰化修饰的细胞壁多糖，使其去乙酰化的过程。
实施例1、BS1蛋白在调控植物细胞壁木聚糖去乙酰化修饰中的应用
一、水稻脆鞘突变体的获得及其BS1蛋白序列分析
1、水稻脆鞘突变体的表型
水稻脆鞘突变体brittle sheath1(简称bs1或突变体bs1)为粳稻品种“黄金晴”的自发突变材料。突变体bs1与粳稻品种“黄金晴”相比，其主要表现为：(1)叶鞘变脆(叶鞘的机械强度显著下降，木质部导管结构异常)；(2)植株变矮。
2、水稻脆鞘突变体的BS1蛋白序列分析
通过测序表明：和野生型水稻植株(黄金晴)相比，水稻脆鞘突变体仅是BS1基因第二个内含子自5’端起第一位碱基由G突变为A，基因组的其余序列与野生型水稻植株全部相同。该碱基突变造成BS1基因第二个内含子未被正常剪切，从而造成BS1蛋白翻译提前终止。
BS1蛋白的氨基酸序列如序列表的序列1所示，BS1基因的核苷酸序列如序列表的序列2所示。BS1蛋白的结构示意图见图1。TM代表跨膜结构域，GDSL代表BS1蛋白主要结构域，AG代表BS1抗血清的抗原片段。
二、转基因植物的制备
1、重组质粒BS1的构建
将序列2所示的DNA分子插入载体pCAMBIA1300的Hind III和Xba I酶切位点之间，且保持载体pCAMBIA1300的其他序列不变，得到重组质粒BS1。
2、转基因植株的获得
将重组质粒进行如下操作：
(1)水稻幼胚培养
将野生型水稻(粳稻品种“黄金晴”)的幼胚脱壳，进行以下消毒流程：依次用70％乙醇溶液表面消毒3min，用0.1％氯化汞溶液浸泡5min，用10％次氯酸钠溶液浸泡20min，之后用无菌水漂洗3-4次。然后播种于NB培养基上以诱导愈伤组织，20天左右从成熟胚盾片处长出愈伤组织，将其继代于NB培养基上。每2周继代一次，每次继代时都挑选色泽淡黄较致密的胚性愈伤组织。
(2)重组农杆菌菌悬液的制备
将重组质粒导入农杆菌EHA105，得到重组农杆菌，用YEP液体培养基培养重组农杆菌至OD600nm为0.8-1.0。
(3)水稻材料与农杆菌的共培养
用步骤(2)制备的重组农杆菌菌悬液侵染步骤(1)制备的胚性愈伤组织，室温振荡浸泡20分钟后取出愈伤组织，在无菌条件下转移到铺有一层无菌滤纸的含20μM乙酰丁香酮的NB固体培养基上，于26℃暗培养2-3天。
(4)抗性愈伤组织的筛选及植株再生
取步骤(3)中经农杆菌侵染的愈伤组织，在含有50mg/L潮霉素的选择培养基上进行筛选培养2周，之后转到第二轮选择培养基上继续筛选2周，选择生长旺盛的抗性愈伤组织转移到分化培养基上分化，再生的小苗在1/2MS固体培养基上生根壮苗，随后移入温室，即为T0代植株。
(5)T0代植株自交，收获种子并培育为植株，即为T1代植株。分别提取T0代植株和T1代植株的基因组DNA，采用抗性基因上的引物(F1：CTTTGAAATTGCCTGATAGA和R1：AAAGTTTGTGGTGTGATTT)进行PCR鉴定，如果待测植株得到大小约为200bp的目的条带，则该植株为阳性转基因植株。
采用重组质粒BS1进行上述步骤，得到40个纯合的转基因株系，随机取其中一个株系进行PCR鉴定，并将鉴定为阳性的转基因株系命名为株系BS1OE。
采用载体pCAMBIA1300进行上述步骤，得到转空载体植株。
三、BS1蛋白在调控植物木聚糖乙酰化修饰水平中的应用
1、乙酰基含量检测
分别取生长4周的突变体bs1、株系BS1OE的T1代植株、转空载体植株的T1代植株和野生型植株(粳稻品种“黄金晴”)，每个株系随机选6株植株，混合并打粉，分别检测待测植物细胞壁总乙酰基含量及分级抽提果胶组分和木聚糖组分的乙酰基含量。具体操作如下：
(1)细胞壁总乙酰基含量检测
将生长4周的植物幼苗冻干至重量不发生变化，将其打磨成粉，并用200目筛筛去粗粒。称取400mg粉末，用20mL 70％乙醇水溶液洗三次，用20mL等体积混合的氯仿-甲醇混合液洗三次，每次漂洗后均以12000rpm离心10min收集沉淀。所得沉淀用丙酮洗涤后烘干，得到以细胞壁成分为主的醇不溶性组分(Alcohol insoluble residue/简称AIR)。用5mLMES/Tris缓冲液(Tris，Sigma，77-86-1；MES，Sigma，1266615-59-1)漂洗沉淀，弃上清。然后加入20mL MES/Tris缓冲液和40U的淀粉酶(Megazyme，K-TDFR-100A)，于97℃反应35min后，转移至60℃水浴锅反应1h，除掉淀粉。2500rpm离心15分钟后，弃上清，加入5mL丙酮漂洗沉淀三次，真空干燥，得到除去淀粉的植物幼苗醇不溶物(AIR)。取6mg除去淀粉的植物幼苗醇不溶物(AIR)溶解于100μL 1N NaOH，28℃200rpm反应1h。再加入100μL 1N HCl中和，12,000rpm离心10分钟，取上清液待测。采用乙酸测定试剂盒(Megazyme，K-ACET)测定上清液中反应释放的乙酸含量(反应释放的乙酸来自于细胞壁中的乙酰基，代表细胞壁中参与反应的一部分乙酰基)。取10μL上清于UV capable 96孔平度板中，并加入94μL水。随后依次加入溶液1和溶液2的混合液42μL(2.5:1)、溶液3和溶液4。分别读取340nm处相应的吸光值A0、A1和A2。利用溶液5来绘制标准曲线，样品中乙酸含量利用如下公式进行计算：
样品＝(A2-A0)-(A1-A0)(A1-A0)/(A2-A0)-Blank；
空白对照＝(A2-A0)-(A1-A0)(A1-A0)/(A2-A0)。
(2)果胶组分的乙酰基含量检测
果胶组分的提取步骤如下：取6mg除去淀粉的植物幼苗醇不溶物(AIR)，于500μL甲酸铵(50mM，pH 4.5)缓冲液中，加入2mU多聚半乳糖醛酸内切酶M2(Megazyme，E-PGALUSP)和0.04mU果胶甲基酯酶(Sigma，9025-98-3)，于37℃反应18h。于80℃孵育20min失活蛋白。3000rpm离心10min，分离上清和沉淀。果胶组分在上清中，木聚糖组分在沉淀中。将上清冻干，即得到果胶组分。将从6mg AIR中提取的全部果胶组分溶解于100μL 1N NaOH，28℃200rpm反应1h。再加入100μL1N HCl中和，12,000rpm离心10分钟，取上清液待测。采用Megazyme的乙酸测定试剂盒测定上清液中反应释放的乙酸含量(反应释放的乙酸来自于细胞壁中的乙酰基，代表细胞壁中参与反应的一部分乙酰基)。具体步骤同步骤(1)。
(3)木聚糖组分的乙酰基含量检测
将从6mg AIR中得到的全部木聚糖组分(步骤(2)中实现了果胶组分和木聚糖组分的分离)溶解于100μL 1N NaOH，28℃200rpm反应1h。再加入100μL 1N HCl中和，12,000rpm离心10分钟，取上清液待测。采用Megazyme的乙酸测定试剂盒测定上清液中反应释放的乙酸含量(反应释放的乙酸来自于细胞壁中的乙酰基，代表细胞壁中参与反应的一部分乙酰基)。具体步骤同步骤(1)。
结果见图2。与野生型植株相比，突变体bs1中的细胞壁总乙酰基含量(释放的乙酸量)显著增加，而株系BS1OE中的细胞壁总乙酰基含量(释放的乙酸量)显著下降。经过分级抽提得到的果胶组分中，三种材料(突变体bs1、株系BS1OE的T1代植株和野生型植株)中的乙酰基含量(释放的乙酸量)无明显差异。而在抽提的木聚糖组分中，突变体bs1中的乙酰基含量(释放的乙酸量)显著高于野生型植株，株系BS1OE中的乙酰基含量(释放的乙酸量)显著下降。
以上结果表明，BS1蛋白具有提高植物细胞壁木聚糖去乙酰化修饰能力的功能，可用于调控植物细胞壁木聚糖的乙酰化修饰水平。
2、BS1蛋白对植物木聚糖乙酰化位点的影响
(1)植物木聚糖的抽提
提取生长4周的突变体bs1和野生型植株(粳稻品种“黄金晴”)的木聚糖。按照步骤1中的(1)方法制备除去淀粉的植物幼苗醇不溶物(AIR)。称取400mg除去淀粉的AIR，加入40mL 1％(质量分数)的草酸铵溶液，于37℃反应过夜，除去果胶组分。将样品离心后收集沉淀，用5mL 11％(体积分数)的过氧乙酸溶液漂洗，弃上清。然后加入20mL 11％(体积分数)的过氧乙酸溶液，85℃反应30分钟。2500rpm离心15分钟，弃上清。加入5mL丙酮漂洗沉淀三次，真空干燥。用20mL的DMSO于70℃过夜进行抽提，重复两次后，将上清转移到新管中。加入5倍体积的乙醇：甲醇：水(7:2:1,v/v,pH 2-3)，于4℃，沉淀3天，得到木聚糖沉淀。2500rpm离心15分钟收集沉淀，再用无水乙醇漂洗三次，真空干燥，即得到木聚糖组分。
(2)利用NMR实验测定木聚糖的乙酰化修饰位点
利用NMR实验测定木聚糖的乙酰化修饰位点，质子共振频率为599.90MHz，1H-NMR和HSQC-NMR实验温度设置为298K，所用探头为5-mm HCN triple resonance的低温探头。Agilent标准脉冲序列gHSQCAD用来检测细胞壁中13C-1H相关单键。所有的1H-13C HSQC谱图的采集范围为：F2(1H)方向谱宽10ppm，F1(13C)谱宽200ppm。采集得到2048×512(F2×F1)的数据矩阵，采样参数：接收增益为30，扫描次数为64次/FID，Interscan delay(d1)为1s。DMSO溶剂峰(dC 39.5ppm和dH 2.49ppm)用来校准波谱。NMR数据的加工和分析使用MestReNova 10.0.2软件。
检测结果如图3所示：突变体bs1中的木聚糖的O-2位和O-3位乙酰化修饰明显低于野生型植株，表明BS1蛋白能特异影响木聚糖O-2和O-3位的乙酰化修饰水平。
序列表
<110> 中国科学院遗传与发育生物学研究所
<120> BS1蛋白在调控植物细胞壁木聚糖乙酰化修饰水平中的应用
<160> 2
<210> 1
<211> 382
<212> PRT
<213> 人工序列
<220>
<223>
<400> 1
Met Gly Ala Val Arg Gly Ile Leu Val Val Ala Val Val Leu Ala Val
1 5 10 15
Ala Ala Ile Leu Ala Gly Ala Ala Glu Gly Lys Val Asn Gly Lys Ala
20 25 30
Lys Gly Lys Tyr Arg Ala Leu Phe Asn Phe Gly Asp Ser Leu Ala Asp
35 40 45
Ala Gly Asn Leu Leu Ala Asn Gly Val Asp Phe Arg Leu Ala Thr Ala
50 55 60
Gln Leu Pro Tyr Gly Gln Thr Phe Pro Gly His Pro Thr Gly Arg Cys
65 70 75 80
Ser Asp Gly Arg Leu Val Val Asp His Leu Ala Asp Glu Phe Gly Leu
85 90 95
Pro Leu Leu Pro Pro Ser Lys Leu Lys Asn Ser Ser Phe Ala His Gly
100 105 110
Ala Asn Phe Ala Ile Thr Gly Ala Thr Ala Leu Asp Thr Pro Tyr Phe
115 120 125
Glu Ala Lys Gly Leu Gly Ala Val Val Trp Asn Ser Gly Ala Leu Leu
130 135 140
Thr Gln Ile Gln Trp Phe Arg Asp Leu Lys Pro Phe Phe Cys Asn Ser
145 150 155 160
Thr Lys Val Glu Cys Asp Glu Phe Tyr Ala Asn Ser Leu Phe Val Val
165 170 175
Gly Glu Phe Gly Gly Asn Asp Tyr Asn Ala Pro Leu Phe Ala Gly Lys
180 185 190
Gly Leu Glu Glu Ala Tyr Lys Phe Met Pro Asp Val Ile Gln Ala Ile
195 200 205
Ser Asp Gly Ile Glu Gln Leu Ile Ala Glu Gly Ala Arg Glu Leu Ile
210 215 220
Val Pro Gly Val Met Pro Thr Gly Cys Phe Pro Val Tyr Leu Asn Met
225 230 235 240
Leu Asp Glu Pro Ala Asp Gly Tyr Gly Pro Gln Ser Gly Cys Val Arg
245 250 255
Arg Tyr Asn Thr Phe Ser Trp Val His Asn Ala His Leu Lys Arg Met
260 265 270
Leu Glu Lys Leu Arg Pro Lys His Pro Asn Val Arg Ile Ile Tyr Gly
275 280 285
Asp Tyr Tyr Thr Pro Val Ile Gln Phe Met Leu Gln Pro Glu Lys Phe
290 295 300
Gly Phe Tyr Lys Gln Leu Pro Arg Ala Cys Cys Gly Ala Pro Gly Ser
305 310 315 320
Val Ala Lys Ala Ala Tyr Asn Phe Asn Val Thr Ala Lys Cys Gly Glu
325 330 335
Ala Gly Ala Thr Ala Cys Asp Asp Pro Ser Thr His Trp Ser Trp Asp
340 345 350
Gly Ile His Leu Thr Glu Ala Ala Tyr Gly His Ile Ala Arg Gly Trp
355 360 365
Val Tyr Gly Pro Phe Ala Asp Gln Pro Ile Phe Gln Ser Ser
370 375 380
<210> 2
<211> 1149bp
<212> DNA
<213> 人工序列
<220>
<223>
<400> 2
atgggggcag ttcgggggat tttggtcgtg gcggtggttc ttgcggtggc ggcgattctt 60
gctggggcgg cggaggggaa ggtgaacggg aaggcgaagg ggaagtacag ggcgctgttc 120
aacttcgggg actcgctggc cgacgccggc aacctcctcg ccaacggcgt cgacttccgc 180
ctcgctaccg cccagctccc ctacggccag accttccccg gccaccccac cggccgctgc 240
tccgacggcc gcctcgtcgt cgaccacctc gccgacgagt tcggcctgcc gctgctgccg 300
ccgtccaagc tcaagaactc cagcttcgct cacggcgcca acttcgccat caccggcgcc 360
accgcgctcg acacccccta cttcgaggcc aaggggctcg gcgccgtcgt ctggaactcc 420
ggcgccctcc tcacccaaat ccagtggttc cgcgatctca agcccttctt ctgcaactcc 480
accaaggtgg aatgcgatga attctatgcg aattcgctct tcgtcgtcgg cgagtttggt 540
ggcaacgact acaatgcgcc gctgtttgcg gggaagggcc ttgaggaggc ctacaagttc 600
atgccggatg tcatccaggc tatctccgat ggcatcgagc aattgattgc tgagggcgca 660
agggagctga ttgtacccgg tgtgatgccc actggatgct tccctgtcta cttgaacatg 720
ctcgatgagc cggccgatgg gtatggcccc cagagcggct gcgtccgtcg gtacaacaca 780
ttctcatggg tgcacaatgc acatctcaag cgcatgcttg agaagctccg gcccaagcac 840
cccaatgtga ggatcatata tggcgattac tacacgcctg ttatccagtt catgcttcag 900
cccgagaagt ttggatttta caagcagcta cctagggcat gctgcggggc tcctgggtcc 960
gttgcgaagg ccgcttacaa cttcaatgtc acagccaaat gtggtgaggc tggtgcaacc 1020
gcgtgtgatg atccatcaac ccattggagc tgggatggca ttcacctgac agaggcggct 1080
tacggtcaca ttgccagagg ttgggtatat ggtcctttcg ctgaccaacc gatcttccaa 1140
tcttcatga 1149
Claims (9)
1.如下a）或b）的蛋白质在调控水稻细胞壁木聚糖乙酰化修饰水平中的应用；
或如下a）或b）的蛋白质在制备调控水稻细胞壁木聚糖乙酰化修饰水平的产品中的应用；
a）氨基酸序列是序列表中序列1所示的蛋白质；
b）在序列表中序列1所示的蛋白质的N端和/或C端连接标签得到的融合蛋白质。
2.与权利要求1中所述的蛋白质相关的生物材料在调控水稻细胞壁木聚糖乙酰化修饰水平中的应用；
或与权利要求1中所述的蛋白质相关的生物材料在制备调控水稻细胞壁木聚糖乙酰化修饰水平的产品中的应用；
所述与权利要求1中所述的蛋白质相关的生物材料为下述A1）至A8）中的任一种：
A1）编码权利要求1中所述的蛋白质的核酸分子；
A2）含有A1）所述核酸分子的表达盒；
A3）含有A1）所述核酸分子的重组载体；
A4）含有A2）所述表达盒的重组载体；
A5）含有A1）所述核酸分子的重组微生物；
A6）含有A2）所述表达盒的重组微生物；
A7）含有A3）所述重组载体的重组微生物；
A8）含有A4）所述重组载体的重组微生物。
3.根据权利要求2所述的应用，其特征在于：A1）所述核酸分子为编码序列是序列表中序列2的DNA分子。
4.根据权利要求1或2所述的应用，其特征在于：所述调控水稻细胞壁木聚糖乙酰化修饰水平为提高水稻细胞壁木聚糖去乙酰化修饰的能力。
5.根据权利要求1或2所述的应用，其特征在于：所述木聚糖乙酰化为木聚糖O-2位和/或O-3位的乙酰化。
6.权利要求1中所述的蛋白质或权利要求2中所述的相关生物材料在培育细胞壁木聚糖乙酰化修饰水平改变的转基因水稻中的应用。
7.一种培育细胞壁木聚糖乙酰化修饰水平降低的转基因水稻的方法，包括在受体水稻中过表达权利要求1中所述的蛋白质，得到转基因水稻的步骤；
所述转基因水稻细胞壁的木聚糖乙酰化修饰水平低于所述受体水稻。
8.根据权利要求7所述的方法，其特征在于：
所述过表达的方法为将权利要求1中所述的蛋白质的编码基因导入受体水稻；
所述转基因水稻细胞壁的木聚糖乙酰化修饰水平低于所述受体水稻体现在转基因水稻细胞壁去乙酰化修饰的能力高于受体水稻。
9.根据权利要求7或8所述的方法，其特征在于：
所述蛋白质的编码基因的核苷酸序列是序列2所示的DNA分子。
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