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作物学报 ›› 2020, Vol. 46 ›› Issue (8): 1217-1224.doi: 10.3724/SP.J.1006.2020.92060

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

OsRPK1基因过表达和RNA干涉对水稻苗期耐盐性的影响

李晶岚,陈鑫欣,石翠翠,刘方惠,孙静,葛荣朝()   

  1. 河北师范大学生命科学学院, 河北石家庄 050024
  • 收稿日期:2019-11-25 接受日期:2020-03-24 出版日期:2020-08-12 网络出版日期:2020-04-21
  • 通讯作者: 葛荣朝
  • 作者简介:李晶岚, E-mail: 83494429@qq.com|陈鑫欣, E-mail: 1046433763@qq.com
  • 基金资助:
    国家自然科学基金项目(30900104);北省自然科学基金项目(C2016205158)

Effects of OsRPK1 gene overexpression and RNAi on the salt-tolerance at seedling stage in rice

LI Jing-Lan,CHEN Xin-Xin,SHI Cui-Cui,LIU Fang-Hui,SUN Jing,GE Rong-Chao()   

  1. College of Life Science, Hebei Normal University, Shijiazhuang 050024, Hebei, China
  • Received:2019-11-25 Accepted:2020-03-24 Published:2020-08-12 Published online:2020-04-21
  • Contact: Rong-Chao GE
  • Supported by:
    National Natural Science Foundation of China(30900104);Hebei Provincial Natural Science Foundation(C2016205158)

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摘要：

水稻OsRPK1基因属于富含亮氨酸重复序列的类受体蛋白激酶, 在盐胁迫下其表达量暂时升高后出现明显下降。前期研究表明, OsRPK1基因过表达会造成拟南芥非生物胁迫耐受性明显降低。本研究对OsRPK1基因在水稻中实现过表达和RNA干涉, 进而对OsRPK1的抗逆性功能加以探究。结果表明, OsRPK1基因表达量增加时, 水稻幼苗表现为耐盐性下降; RNA干涉则使水稻幼苗表现为耐盐性增加。盐胁迫后OsRPK1过表达植株脯氨酸含量低于野生型、MDA含量和相对电导率显著高于野生型, 而OsRPK1-RNAi水稻植株的脯氨酸含量显著高于野生型、MDA含量和相对电导率显著低于野生型。因此, OsRPK1基因的表达量对水稻耐盐性具有明显影响, 脯氨酸含量及细胞质膜受破损程度的改变可能是造成转基因水稻耐盐性变化的内在原因。本研究为进一步阐明OsRPK1基因的抗逆作用机制奠定了基础, 对于通过调整该基因表达改良水稻的耐盐性具有重要意义。

关键词: 水稻, OsRPK1基因, 过表达, RNA干涉, 生理指标

Abstract:

OsRPK1 is a leucine-rich repeat receptor-like protein kinase (LRR-RLK) gene in rice, and its expression level decreases evidently after temporarily increasing under salt stress. Previous studies have shown that the overexpression of OsRPK1 can significantly reduce the abiotic stress tolerance of Arabidopsis. In this study, OsRPK1 gene was overexpressed and RNA interfered in rice. The salt tolerance of the 35S:OsRPK1 rice seedlings was significantly lower than that of the control plants. The OsRPK1-RNAi rice seedlings exhibited higher salt tolerance than the wild-type plants. The proline content of OsRPK1 overexpressed plants was lower than that of the wild type, MDA content and relative conductivity were significantly higher than those of the wild type, while the proline content of OsRPK1-RNAi rice plants was significantly higher than those of wild type, MDA content and relative conductivity were significantly lower than those of wild type. The results of this study indicate that OsRPK1 plays an important role in salt tolerance of rice. The changes of proline content and the damage degree of cytoplasmic membrane may be the internal reasons for the salt-tolerance variations of the transgenic rice lines. This study lays a foundation for further elucidating the resistance mechanism of OsRPK1 gene, which is very important for cultivating salt-tolerance rice by adjusting the expression of OsRPK1 gene.

Key words: rice, OsRPK1 genes, overexpressed, RNA interference, physiological indexes

图1

OsRPK1基因在140 mmol L-1 NaCl胁迫的水稻中表达量的变化 差异显著性分析采用t-test方法, *P < 0.05、**P < 0.01、***P < 0.001。"

图2

35S:OsRPK1植株与OsRPK1-RNAi植株的qRT-PCR检测 差异显著性分析采用t-test方法, *P < 0.05、**P < 0.01、 ***P < 0.001。"

图3

OsRPK1过表达水稻在盐胁迫下的生长情况 A: NaCl胁迫前; B: 140 mmol L-1 NaCl胁迫6 d; C: 140 mmol L-1 NaCl胁迫后恢复9 d; D: 盐胁迫恢复培养后存活率统计。差异显著性分析采用t-test方法, *P < 0.05、**P < 0.01、***P < 0.001。"

图4

OsRPK1-RNAi水稻在胁迫下生长情况 A: NaCl胁迫前; B: 140 mmol L-1 NaCl胁迫10 d; C: 140 mmol L-1 NaCl胁迫后恢复7 d; D, E: 盐胁迫恢复培养后存活率统计。差异显著性分析采用t-test方法, *P < 0.05、**P < 0.01、***P < 0.001。"

图5

140 mmol L-1 NaCl盐胁迫后转基因水稻脯氨酸含量 差异显著性分析采用t-test方法, *P < 0.05、**P < 0.01、***P < 0.001。"

图6

140 mmol L-1 NaCl盐胁迫后转基因水稻株系中MDA含量 差异显著性分析采用t-test方法, *P < 0.05、**P < 0.01、***P<0.001。"

图7

转基因水稻的相对电导率 差异显著性分析采用t-test方法, *P < 0.05、**P < 0.01、***P < 0.001。"

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