CN114805522B - 水稻OsbHLH38蛋白及其编码基因在提高植物抗非生物胁迫中的用途 - Google Patents
水稻OsbHLH38蛋白及其编码基因在提高植物抗非生物胁迫中的用途 Download PDFInfo
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Abstract
本发明公开了水稻OsbHLH38蛋白及其编码基因在提高植物抗非生物胁迫中的用途。本发明分别采用超表达和CRISPR敲除技术将水稻OsbHLH38基因在水稻中进行超表达或进行敲除突变，根据转基因水稻植株的表型变化发现，在水稻中超表达OsbHLH38基因能够显著改善水稻抵御盐胁迫的能力；因此，本发明提供了水稻OsbHLH38蛋白及其编码基因能够在提高植物对于非生物胁迫抗性中的应用，包括：将水稻OsbHLH38基因在植物中进行过表达得到转基因植物，所得到的转基因植物对于非生物胁迫的抗性明显增强。本发明在改良、增强水稻抗逆性以及加速抗逆分子育种进程等方面具有应用前景。
Description
技术领域
本发明涉及水稻OsbHLH38蛋白及其编码基因的新用途，尤其涉及水稻OsbHLH38蛋白及其编码基因在提高植物抗非生物胁迫中的新用途，属于水稻OsbHLH38蛋白及其编码基因的新用途领域。
背景技术
干旱和盐碱等非生物胁迫对水稻(Oryza sativa L)生产带来不利影响，并对粮食安全造成严重威胁。仅靠传统的作物育种技术不足以实现作物对干旱胁迫的耐受性。近年来，在水稻抗逆耐盐基因功能解析和分子育种、等方面取得了一系列重大进展，对抗逆分子调控机制的解析仍是当前逆境研究的重要课题。
本发明人课题组前期通过蛋白互作发现OsbHLH38(Basic helix-loop-helix 38）除了可以与已验证的抗逆侯基因OsDREB2A，OsMAR1相互作用，并且OsbHLH38受ABA(Abscisic Acid)诱导上调表达。ABA是一种植物激素，植物通过利用它来协调对非生物胁迫的响应、调节种子休眠(Mehrotra R, Bhalothia P, Bansal P, et al. Abscisic acidand abiotic stress tolerance–Different tiers of regulation. Journal of plantphysiology, 2014, 171(7): 486-496.)。有研究表明体外添加ABA阻断剂ANT(Antabactin)能够使处于休眠的种子恢复活力，促进种子萌发率(Vaidya A S, PetersonF C, Eckhardt J, et al. Click-to-lead design of a picomolar ABA receptorantagonist with potent activity in vivo. Proceedings of the National Academyof Sciences, 2021, 118(38).)。在拟南芥中OsbHLH38的同源基因AtbHLH38被报道与AtbHLH39协同参与调控铁离子吸收(Yuan Y, Wu H, Wang N, et al. FIT interactswith AtbHLH38 and AtbHLH39 in regulating iron uptake gene expression for ironhomeostasis in Arabidopsis. Cell research, 2008, 18(3): 385-397)。迄今为止，未见水稻中OsbHLH38蛋白或其编码基因在提高植物非生物胁迫作用的任何报道。
发明内容
本发明的主要目的在于提供水稻OsbHLH38蛋白或其编码基因在提高植物抗非生物胁迫中的用途。
为了实现上述之发明目的，本发明主要采取的技术方案包括：
本发明的一方面是提供水稻OsbHLH38蛋白或其编码基因在提高植物抗非生物胁迫中的用途；优选的，所述的非生物胁迫包括盐胁迫或渗透胁迫。
本发明的第二方面是提供了一种提高植物对非生物胁迫抗性的方法，包括：将水稻OsbHLH38基因在植物中进行过表达得到转基因植物；所得到的转基因植物对于非生物胁迫的抗性增强；譬如：将水稻OsbHLH38基因可操作的与表达调控元件相连接，得到在植物中表达该编码基因的重组植物表达载体；将所述重组植物表达载体转化受体植物，使水稻OsbHLH38基因在植物中进行过表达；优选的，所述的非生物胁迫包括盐胁迫或渗透胁迫。
作为本发明一种优选的具体实施方案，所述的提高植物对非生物胁迫抗性的具体方法包括：（1）构建含有水稻OsbHLH38基因的重组植物表达载体；（2）将所构建的重组植物表达载体转化到受体植物组织或植物细胞中；（3）培育筛选得到对非生物胁迫抗性提高的转基因植物。
本发明的第三方面是提供了一种培育抗非生物胁迫的植物新品种的方法，包括：（1）构建含有水稻OsbHLH38基因的重组植物表达载体；（2）将所构建的重组植物表达载体转化到受体植物组织或植物细胞中；（3）培育筛选得到对非生物胁迫抗性提高的植物新品种；优选的，所述的非生物胁迫包括盐胁迫或渗透胁迫。
本发明进一步提供了含有所述水稻OsbHLH38基因的重组植物表达载体以及含有所述重组植物表达载体的重组宿主细胞；将所述水稻OsbHLH38基因可操作的与表达调控元件相连接得到重组植物表达载体；该重组植物表达载体可以由5′端非编码区，水稻OsbHLH38基因和3′非编码区组成；其中，所述的5′端非编码区可以包括启动子序列、增强子序列或/和翻译增强序列；所述的启动子可以是组成性启动子、诱导型启动子、增强型启动子、组织或器官特异性启动子；所述的3′非编码区可以包含终止子序列、mRNA切割序列等。合适的终止子序列可取自根癌农杆菌的Ti-质粒，例如章鱼碱合成酶和胭脂碱合成酶终止区。
所述重组植物表达载体还可含有用于选择转化细胞的选择性标记基因，用于选择经转化的细胞或组织。所述标记基因包括：编码抗生素抗性的基因以及赋予除草化合物抗性的基因等。此外，所述的标记基因还包括表型标记，例如β-半乳糖苷酶和荧光蛋白等。
作为一种参考的具体实施方案，所述重组植物表达载体构建包括：在植物表达载体pMDC43的GFP下游attR1与attR2之间插入水稻OsbHLH38基因。
转化方案以及将所述基因（或多核苷酸）引入植物的方案可视用于转化的植物（单子叶植物或双子叶植物）或植物细胞的类型而变化。将所述基因多（或核苷酸）引入植物细胞的合适方法包括：显微注射、电穿孔、农杆菌介导的转化、直接基因转移以及高速弹道轰击等。在特定的实施方案中，可利用多种瞬时转化法将所述基因（或多核苷酸）提供给植物。利用常规方法可使已转化的细胞再生稳定转化植株。
本发明可用于转化任何植物种类，所述的植物包括但不限于单子叶植物或双子叶植物；更优选的，所述的植物包括农作物、蔬菜或观赏植物、果树等，例如，可以是水稻、棉花、玉米、高粱、小麦、大豆、马铃薯、大麦、番茄、菜豆、花生或甘蔗等，优选为水稻。
作为本发明一种优选的具体实施方案，所述的非生物胁迫包括盐胁迫或渗透胁迫。
作为本发明一种优选的具体实施方案，所述水稻OsbHLH38蛋白的氨基酸序列为SEQ ID NO.2所示；所述水稻OsbHLH38蛋白的编码基因的CDS核苷酸序列为SEQ ID NO.1所示。另外，本领域技术人员还可以将SEQ ID NO.1所示的多核苷酸进行优化以增强在植物中（尤其是水稻）的表达效率。
本发明人前期通过蛋白互作发现OsbHLH38可以与已验证的抗逆侯基因OsDREB2A，OsMAR1相互作用，并且OsDREB2A受ABA诱导上调表达。本发明进一步通过120mM NaCl条件下，超表达和CRISPR敲除水稻植株的表型变化，进行其基因克隆与功能分析，分析候选基因与水稻非生物胁迫应答的关系，结果表明在水稻中超表达OsbHLH38基因能够显著改善水稻抵御盐胁迫或渗透胁迫的能力。本发明为改良、增强水稻抗逆性，加速抗逆分子育种进程，具有十分重要的理论和实际意义。
本发明所涉及到的术语定义
除非另外定义，否则本文所用的所有技术及科学术语都具有与本发明所属领域的普通技术人员通常所了解相同的含义。
术语“多核苷酸”或“核苷酸”意指单股或双股形式的脱氧核糖核苷酸、脱氧核糖核苷、核糖核苷或核糖核苷酸及其聚合物。除非特定限制，否则所述术语涵盖含有天然核苷酸的已知类似物的核酸，所述类似物具有类似于参考核酸的结合特性并以类似于天然产生的核苷酸的方式进行代谢。除非另外特定限制，否则所述术语也意指寡核苷酸类似物，其包括PNA（肽核酸）、在反义技术中所用的DNA类似物（硫代磷酸酯、磷酰胺酸酯等）。除非另外指定，否则特定核酸序列也隐含地涵盖其保守修饰的变异体（包括（但不限于）简并密码子取代）和互补序列以及明确指定的序列。特定而言，可通过产生其中一个或一个以上所选（或所有）密码子的第3位经混合碱基和/或脱氧肌苷残基取代的序列来实现简并密码子取代。
术语“多肽”、“肽”和“蛋白”在本文中互换使用以意指氨基酸残基的聚合物。即，针对多肽的描述同样适用于描述肽和描述蛋白，且反之亦然。所述术语适用于天然产生氨基酸聚合物以及其中一个或一个以上氨基酸残基为非天然编码氨基酸的氨基酸聚合物。如本文中所使用，所述术语涵盖任何长度的氨基酸链，其包括全长蛋白(即抗原)，其中氨基酸残基经由共价肽键连接。
术语“重组宿主细胞株”或“宿主细胞”意指包含本发明多核苷酸的细胞，而不管使用何种方法进行插入以产生重组宿主细胞，例如直接摄取、转导、f配对或所属领域中已知的其它方法。外源性多核苷酸可保持为例如质粒的非整合载体或者可整合入宿主基因组中。宿主细胞可为原核细胞或真核细胞，宿主细胞还可为单子叶或双子叶植物细胞。
术语“可操作的连接”指两个或更多个元件之间功能性的连接，可操作的连接的元件可为邻接或非邻接的。
术语“重组植物表达载体”意指一种或多种用于实现植物转化的DNA载体；本领域中这些载体常被称为二元载体。二元载体连同具有辅助质粒的载体是大多常用于土壤杆菌介导转化的。二元载体通常包括：T-DNA转移所需要的顺式作用序列、经工程化处理以便能够在植物细胞中表达的选择标记物，待转录的异源性DNA序列等。
本发明中所述术语“转化”指将水稻OsbHLH38基因导入到植物细胞内部这样的方式将多核苷酸或多肽遗传转化到植物中。将所述多核苷酸或多肽引入到植物中的方法为本领域所习知，包括但不限于稳定转化法、瞬时转化法和病毒介导法等。“稳定转化”指被引入的多核苷酸构建体整合至植物细胞的基因组中并能通过其子代遗传；“瞬时转化”指多核苷酸被引入到植物中但只能在植物中暂时性表达或存在。
附图说明
图1为过表达OsbHLH38转基因植株PCR阳性鉴定，其中，1-9号泳道为转基因材料阳性植株，10号(+)泳道为质粒载体作为阳性对照，11号(-)泳道为野生型阴性对照。
图2为水稻OsbHLH38基因的不同株系的敲除情况，其中，KO-6株系为55 bp的大片段缺失，KO-9株系为2 bp的双碱基缺失。
图3为水稻OsbHLH38基因过表达和敲除转基因材料的各个株系的表达量情况qRT-PCR检测结果。
图4为水稻OsbHLH38基因过表达、CRISPR-Cas9敲除转基因植株与野生型植株在120 umol NaCl胁迫下的表型；上半部分为NaCl胁迫后7天的表型图片，下半部分为NaCl胁迫后恢复培养5天后的表型图片，WT为野生型，KO为敲除材料，OE为过表达材料。
图5为水稻OsbHLH38基因过表达、CRISPR-Cas9敲除转基因植株与野生型植株在120 umol NaCl胁迫后7天进行复水后存活率统计图片，WT为野生型，KO为敲除材料，OE为过表达材料。
具体实施方式
以下结合具体实施例来进一步描述本发明，本发明的优点和特点将会随着描述而更为清楚。但这些实施例仅是范例性的，并不对本发明的范围构成任何限制。本领域技术人员应该理解的是，在不偏离本发明的精神和范围下可以对本发明的细节和形式进行修改或替换，但这些修改和替换均落入本发明的保护范围内。
实施例1 OsbHLH38基因在水稻中的遗传转化实验
⒈OsbHLH38基因过表达载体构建
首先用用限制性内切酶EmaI1051消化pGWC载体(公众可从中国农业科学院作物科学研究所获得该载体) ，使其线性化并回收。
根据日本晴序列全长设计引物，以日本晴cDNA为模板，扩增获得 OsbHLH38蛋白的编码区全长序列，同时在5 ′和3 ′端加上pGWC载体线性化后的接头，扩增引物为：
PCR扩增并回收目的DNA片段(1068bp)。采用In-Fusion HD Cloning Kit(Clontech，Code no：639648)进行目的片段与线性化载体的同源重组，阳性克隆质粒进行PCR和测序验证，测序结果表明在载体pGWC的两个EmaI1051酶切位点之间插入了SEQ IDNO.1所示的OsbHLH38基因片段（该序列所编码OsbHLH38蛋白的氨基酸序列为SEQ ID NO .2所示）得到入门载体，将重组载体命名为pGWC-OsbHLH38。
采用
LR
II enzyme mix(Invitrogen，Code no：11791020)进行最终载体构建，阳性克隆质粒进行PCR和测序验证，测序结果表明在载体pMDC43(公众可从中国农业科学院作物科学研究所获得该载体)的GFP下游插入了SEQ ID NO .1所示的OsbHLH38基因片段，该序列所编码的OsbHLH38蛋白的氨基酸序列如SEQ ID NO .2所示，将重组载体命名为pMDC43-OsbHLH38。
⒉OsbHLH38基因CRISPR-Cas9敲除载体构建
根据OsbHLH38基因cDNA序列设计敲除的靶位点，所用网站为：http://skl.scau.edu.cn/sadsdecode/；
靶位点序列：5 ′- CCCTTCTACTTCGACGGTAGGCGG-3 ′，
实验方法及载体来自刘耀光院士实验室(Ma X ,Zhang Q ,Zhu Q ,Liu W ,ChenY,Qiu R , Wang B ,Yang Z ,Li H ,Lin Y ,Xie Y , Shen R ,Chen S ,Wang Z ,Chen Y,Guo J , Chen L ,Zhao X ,Dong Z ,Liu YG .A Robust CRISPR/Cas9 System forConvenient ,High-Efficiency Multiplex Genome Editing in Monocot and DicotPlants .Molecular Plant ,2015 ,8(8):1274-1284) ，将重组载体命名为 pYLCRISPR/Cas9Pubi-OsbHLH38。
⒊农杆菌转化
冻融法将表达载体pMDC43-OsbHLH38和敲除载体 pYLCRISPR/Cas9Pubi-OsbHLH38转入农杆菌EHA105感受态细胞中(公众可从中国农业科学院作物科学研究所获得，记载过该材料的非专利文献是：Ruifang Yang ,Qicai Tang ,HuimeiWang ,Xiaobo Zhang ,GangPan ,HongWang and Jumin Tu Analyses of two rice(Oryza sativa)cyclin-dependentkinase inhibitors and effects of transgenic expression of OsiICK6 on plantgrowth and development，2011,Annals of Botany ,107:1087-1101) ，方法参照分子克隆实验指南。
⒋遗传转化
用农杆菌介导的遗传转化法，以日本晴为受体材料，进行遗传转化，培养基配方见表1。
表1 遗传转化所用培养基及其配方
具体方法如下：
⑴愈伤诱导
取适量成熟的水稻种子，脱壳后，先用70％酒精清洗消毒1min，期间不断地摇荡，再用15％的次氯酸钠消毒处理30min(可置于摇床上振荡)；最后用无菌蒸馏水冲洗4～5次，用无菌滤纸吸干种子表面水分后接种。将消毒处理后的种子接种于含有2.0mg/L的2，4-D的诱导培养基中，28℃暗培养30～40天，培养获得的愈伤在继代培养基上扩大培养，每2周继代一次，直至胚性愈伤形成。
⑵农杆菌侵染
a)将携带有表达质粒载体pMDC43-OsbHLH38和 pYLCRISPR/Cas9Pubi- OsbHLH38的农杆菌划线于含有抗生素(50mg/L卡那霉素或壮观霉素、25mg/L利福平)的LB固体培养基表面，28℃，200rpm 培养过夜。
b)用灭过菌的牙签挑取单克隆菌落，接种到5mL含有相应抗生素的YEB液体培养基中，28℃震荡培养到OD600=0 .5。
c)取活化好的的新鲜菌液按1:100的比例接种到25mL相同的YEB液体培养基中，在相同条件下培养至OD600=0 .5。
d)菌液在5000g，4℃下离心10min收集菌体，弃去上清液；加入25mL 10mM的MgSO4悬浮菌体，并用移液枪轻轻吸打使其充分悬浮，在5000g，4℃下离心10min重新收集菌体，弃去上清液。
e)用25mL含有200μM乙酰丁香酮(AS)的AA-AS浸染培养基重新悬浮。
f)将生长状态良好的胚性愈伤从继代培养基中转移至表面覆有无菌滤纸的培养皿中(愈伤切成0 .3～0.4mm大小) ，在超净工作台上风干10～20min。
g)将干燥的胚性愈伤浸入含有上述菌液的50mL离心管中20min，期间每隔5min摇荡一次；之后倒掉菌液，将愈伤取出置于无菌滤纸上风干10～20 min，然后转移至表面铺有无菌滤纸的含有200μM乙酰丁香酮(AS)的CC培养基中，25℃黑暗条件下共培养3天。
h)收集表面没有明显农杆菌的愈伤组织，用含600mg/L头孢霉素的无菌水冲洗3次，吸取多余水分。
i)将愈伤组织转入筛选培养基(含500mg/L头孢霉素和50mg/L潮霉素的N6培养基)上继续筛选2～3次，每次两周。最终得到生长良好的鲜黄色潮霉素抗性愈伤组织。
⑶转化体植株再生
取新鲜潮霉素抗性的愈伤，将愈伤组织分割成2mm的小块，接于预分化培养基中，28℃暗培养7天，然后置于光照培养间(12h光照/12h黑暗)继续培养8-9天后将已分化出不定芽的愈伤组织后转入再生培养基(250mL组培瓶) 上，继续光照培养。等到不定芽长成4～6cm高的小苗后再转移到生根培养基中，在28℃光照培养间(12h光照/12h黑暗)条件下培养约15天，得到转化体植株，移到温室种植(T0代)，一月后取叶片进行PCR阳性鉴定(Hpt-F：5’-CTATTTCTTTGCCCTCGGAC-3’，Hpt-R：5’-CCTGACCTATTGCATCTCCC-3’) ，将鉴定为阳性的转基因植物收获其阳性植株种子 (T1代)。
以下OE及KO引物是用于后续材料鉴定是否纯合使用的鉴定引物：
OE引物：
正向引物为OE-F: 5'-ATGGCATGGATGAACTATA -3'
反向引物为OE-R: 5'-CCAGCCACTAGAATTGAAG -3'
KO引物：
正向引物为KO-F: TGGTTTGGTTGTTGTGGTGG
反向引物为KO-R: GCACTGTGAGATTGTGAGAAG
图1为过表达材料敲除材料转基因植株PCR鉴定结果。
图2为不同敲除类型与野生型的基因序列比对。
⒌转基因植株分子鉴定和抗逆性鉴定
选用T2代OsbHLH38转基因超表达、OsbHLH38基因CRISPR-Cas9纯和种子及野生型日本晴的种子，发芽后播种于盛有草炭土的盒子内，培养条件为光照/黑暗为16/8h，光照条件26℃，黑暗条件22℃，光强30000lx。利用荧光定量PCR检测转基因水稻在RNA水平的表达量。
图3为OsbHLH38基因过表达材料和敲除材料的不同株系的表达量。从图3中可见，相比于野生型材料，OsbHLH38基因过表达材料中OsbHLH38表达量显著上升，而敲除材料中OsbHLH38基因表达量显著下降。
在水稻生长至三叶期用120 umol NaCl处理，每胁迫处理进行6次随机重复，实验设3次重复，试验结果见图4和图5。图4中左部是OsbHLH38转基因敲除材料与野生型植株NaCl在胁迫后7天和NaCl胁迫后恢复培养5天后的表型，右部OsbHLH38转基因过表达材料与野生型植株在NaCl胁迫后7天和NaCl胁迫后恢复培养5天后的表型；图5为水稻OsbHLH38基因过表达、CRISPR-Cas9敲除转基因植株与野生型植株在120 umol NaCl胁迫后7天进行复水后存活率统计图片。
根据图4和图5的实验结果可见，在水稻中超表达水稻OsbHLH38基因能够显著改善水稻抵御盐胁迫的能力，盐胁迫复水后存活率较野生型显著提高。将水稻中的OsbHLH38基因进行敲除后会明显降低水稻抵御盐胁迫的能力，盐胁迫复水后存活率较野生型显著下降。因此，水稻OsbHLH38蛋白及其编码基因和重组载体能够应用于增强作物的抗盐胁迫能力。
序列表
<110> 中国农业科学院作物科学研究所
<120> 水稻OsbHLH38蛋白及其编码基因在提高植物抗非生物胁迫中的用途
<130> BJ-2011-220613A-L
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1038
<212> DNA
<213> Oryza sativa L
<400> 1
atgtgggaag ctgtaggcgg cggcgacggc acggctctcc tgccgtggcc cggctcggcc 60
gcggccgcgc cgctctacat gcccccggca gcagcggcag cggcgccctt cgccgccggc 120
gagcagctgc cggtggagca gcccttctac ttcgacggag gcggaggcgt ggcggggcat 180
aatcaccatc ctcatcatca tcagtacggg atggaggcgc cgccgccgat gacgatgatg 240
cagatgggcg gcgggggttc gtcgtcgtcg aggatggtgg tgtccgggct gctcgggacg 300
ctgcaggcgg agctcgggag gatgacggcg aaggagatca tggacgccaa ggcgctggcg 360
gcgtcgcgca gccacagcga ggccgagcgc cgccgccgcc agcgcatcaa cggccacctc 420
gccaggctcc gcagcctcct ccccaacacc accaagacgg acaaggcgtc gctgctggcg 480
gaggtgatcg agcacgtcaa ggagctgaag cggcagacgt cggcgatgat ggaggacggc 540
gccgcgggag gggaggcggc ggcggcgccg gtggtgctgc tgccgacgga ggacgacgag 600
ctggaggtgg acgcggcggc ggacgaggga gggaggctcg tggcgagggc ctcgctgtgc 660
tgcgaggacc gcgccgacct catcccgggc atcgcccgcg cgctcgccgc gctccggctg 720
cgcgcccgcc gcgccgagat cgccacgctc ggcggccgcg tccgcagcgt cctcctcatc 780
gccgccgtcg aggaggaaga tccagacgag gccgggaacg acgacgacgg cgaacacggc 840
tacggcgttg ccgcctcgca ccggaggcac gagctcgtcg cgtcgatcca cgaggcgctg 900
cgcggcgtca tgaaccgcaa agcggcgagc agcgacacct cctcttccgg cgccggcggc 960
ggcggtggga gcatcaagag gcagcggatg atcagtgcac atgatcagca aggttccttc 1020
aattctagtg gctggtaa 1038
<210> 2
<211> 345
<212> PRT
<213> Oryza sativa L
<400> 2
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His His His Gln Tyr Gly Met Glu Ala Pro Pro Pro Met Thr Met Met
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Glu Val Ile Glu His Val Lys Glu Leu Lys Arg Gln Thr Ser Ala Met
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Claims (4)
1. 水稻OsbHLH38蛋白或水稻OsbHLH38蛋白的编码基因在提高植物对于非生物胁迫抗性中的用途；所述的非生物胁迫包括盐胁迫或渗透胁迫；所述的植物是水稻；所述水稻OsbHLH38蛋白的氨基酸序列为SEQ ID NO.2所示；所述水稻OsbHLH38蛋白的编码基因的CDS的核苷酸序列为SEQ ID NO.1所示。
2.一种提高植物对非生物胁迫抗性的方法，其特征在于，包括：将水稻OsbHLH38基因的CDS在植物中进行过表达得到转基因植物；所述水稻OsbHLH38基因的CDS的核苷酸序列为SEQ ID NO.1所示；所述非生物胁迫包括盐胁迫或渗透胁迫；所述的植物是水稻。
3.根据权利要求2所述的方法，其特征在于，包括：（1）构建含有水稻OsbHLH38基因的CDS的重组植物表达载体；（2）将所构建的重组植物表达载体转化到植物组织或植物细胞中；（3）培育筛选得到对非生物胁迫抗性提高的转基因植物。
4.一种培育抗非生物胁迫的植物新品种的方法，其特征在于，包括：（1）构建含有水稻OsbHLH38基因的CDS的重组植物表达载体；所述水稻OsbHLH38基因的CDS的核苷酸序列为SEQ ID NO.1所示；（2）将所构建的重组植物表达载体转化到植物组织或植物细胞中；（3）培育筛选得到对非生物胁迫抗性提高的植物新品种；所述非生物胁迫包括盐胁迫或渗透胁迫；所述的植物是水稻。
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