水稻OsCNGC10基因抗倒伏性以及抗旱性功能研究
Mechanism of loding residence and drought tolerance of OsCNGC10 gene in rice
朱忠林 1文月 1周棋 1巫燕飞 1杜雪竹 2盛锋1
作者信息
- 1. 湖北大学生命科学学院 / 省部共建生物催化与酶工程国家重点实验室, 湖北武汉 430062
- 2. 湖北大学生命科学学院 / 省部共建生物催化与酶工程国家重点实验室, 湖北武汉 430062;湖北洪山实验室, 湖北武汉 430062
- 折叠
摘要
环核苷酸门控离子通道是一种配体门控的阳离子通道,存在于动物和植物体内,是真核生物信号级联反应的重要组成部分.本研究利用水稻环核苷酸门控离子通道(cyclic nucleotide-gated channel,OsCNGC10)基因,构建了超表达载体 pU1301-CNGC10-Flag和双靶点敲除载体pRGEB32-CRISPR/cas9-cngc10,通过农杆菌介导的遗传转化法获得敲除和过表达材料,并从 T2 代中分离到纯合植株 oscngc10-2 及 OE-CNGC10-6.转基因植株茎秆特性以及抗倒伏性分析表明,突变体 oscngc10-2 茎秆强度和抗倒伏性增强;茎秆细胞壁组织切片以及组织成分分析则表明突变体oscngc10-2 植株抗倒伏性增强是由于茎秆细胞壁茎壁厚度、薄壁组织细胞丰度以及木质素含量增加所致;过表达OsCNGC10 降低了茎秆壁厚、茎秆木质素含量以及茎秆细胞壁细胞丰度,敲除OsCNGC10 增加了茎秆木质素含量且增加了茎秆细胞壁薄壁细胞丰度,初步证明 OsCNGC10 与水稻茎秆细胞壁成分合成相关,负调控水稻抗倒伏性;T2代田间试验结果表明,与野生型相比,突变体 oscngc10-2 植株的株高、有效穗、穗长、穗粒数、结实率、千粒重和单株产量等农艺性状显著提升;苗期干旱胁迫实验结果表明,在干旱胁迫下,OsCNGC10基因缺陷型植株体内丙二醛(MDA)含量积累速度加快,且无法形成足够的游离脯氨酸,而过表达 OsCNGC10 植株在遭受干旱胁迫时,体内游离脯氨酸(Pro)含量大量升高,且 MDA 积累速度相对变慢,初步说明 OsCNGC10 正向调控水稻苗期抗旱性.本研究结果表明水稻OsCNGC10 可能在水稻抗倒伏及抗旱方面有潜在功能,为培育抗倒伏且高产的水稻新品种提供了理论基础和新的种质资源.
Abstract
Cyclic nucleotide-gated ion channels are ligand-gated cationic channels that exist in animals and plants,which are an important part of eukaryotic signaling cascades.In this study,OsCNGC10(cyclic nucleotide-gated channel)gene in rice was used,and the overexpression vector pU1301-CNGC10-Flag and the double-target knockout vector pRGEB32-CRISPR/cas9-cngc10 were constructed.The knockout and overexpression materials were obtained by Agrobacterium-mediated genetic transformation.Homozygous plants oscngc10-2 and OE-CNGC10-6 were isolated from T2 generation.The analysis of stem characteristics and lodging resistance of transgenic plants showed that oscngc10-2 had enhanced stem strength and lodging resistance.Stem cell wall sections and tissue composition analysis showed that oscngc10-2 increased lodging resistance due to the increase of stem wall thickness,parenchyma cell abundance,and lignin content.The knockout of OsCNGC10 increased the lignin content and the abundance of stem-cell wall parenchyma cells.The overexpression of OsCNGC10 reduced stem wall thickness,lignin content,and cell abundance in stem cell wall,while the knockdown of OsCNGC10 increased lignin content and increased the abundance of thin-walled cells in stem cell wall,suggesting that OsCNGC10 was associated with the composition of stem cell wall and negatively regulated lodging resistance in rice.T2 generation field experiment indicated that compared with the wild type,oscngc10-2 significantly increased plant height,the effective panicle length,panicle number,seed setting rate,1000-grain weight,and yield per plant.The results of drought stress at seedling stage showed that malondialdehyde(MDA)content accumulated rap-idly in OsCNGC10 defective plants under drought stress and insufficient free proline(Pro)was formed,while the free Pro content in OsCNGC10 plants was significantly increased.Moreover,the MDA accumulation rate was relatively slow,which preliminarily indicated that OsCNGC10 positively regulated the drought resistance at seedling stage.The results of this study indicated that OsCNGC10 might have a potential function in lodging resistance and drought resistance in rice,which providing a theoretical basis and new germplasm resources for the breeding lodging resistance and high yield of new rice varieties.
关键词
水稻/OsCNGC10/抗倒伏/抗旱/细胞壁Key words
rice/OsCNGC10/lodging resistance/drought tolerance/cell wall引用本文复制引用
基金项目
武汉市生物技术关键技术科技重大专项(2022021302024851)
粮食作物种质创新与遗传改良湖北省重点实验室开放基金(2018lzjj01)
出版年
2024
国家科技期刊平台
NSTL
万方数据