水稻OsPR10A的表达特征及其在干旱胁迫应答过程中的功能

doi:10.11983/CBB19042

植物学报 ›› 2019, Vol. 54 ›› Issue (6): 711-722.DOI: 10.11983/CBB19042

水稻OsPR10A的表达特征及其在干旱胁迫应答过程中的功能

张彤¹,郭亚璐^1,²,陈悦¹,马金姣¹,兰金苹^1,³,燕高伟¹,刘玉晴¹,徐珊¹,李莉云¹,刘国振^1,^*(),窦世娟^1,^*()

1 河北农业大学生命科学学院, 保定 071001
2 中国农业科学院农业基因组研究所, 深圳 518116
3 河北北方学院生命科学研究中心, 张家口 075000

收稿日期:2019-02-27 接受日期:2019-05-06 出版日期:2019-11-01 发布日期:2020-07-09
通讯作者: 刘国振,窦世娟
基金资助:
国家自然科学基金(31400700)

Expression Characterization of Rice OsPR10A and Its Function in Response to Drought Stress

Tong Zhang¹,Yalu Guo^1,²,Yue Chen¹,Jinjiao Ma¹,Jinping Lan^1,³,Gaowei Yan¹,Yuqing Liu¹,Shan Xu¹,Liyun Li¹,Guozhen Liu^1,^*(),Shijuan Dou^1,^*()

1 College of Life Sciences, Hebei Agricultural University, Baoding 071001, China
2 Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518116, China
3 Research Center for Life Sciences, Hebei North University, Zhangjiakou 075000, China

Received:2019-02-27 Accepted:2019-05-06 Online:2019-11-01 Published:2020-07-09
Contact: Guozhen Liu,Shijuan Dou

摘要/Abstract

摘要： 利用免疫印迹(WB)分析了水稻(Oryza sativa) OsPR10A在其不同生长时期、不同组织部位及多种非生物逆境胁迫下的表达特征, 发现OsPR10A在干旱、盐胁迫以及茉莉酸甲酯(MeJA)和脱落酸(ABA)诱导下表达量明显升高, 表明该蛋白可能在干旱和盐胁迫应答过程中发挥作用。为证明这一推测, 我们构建了OsPR10A超表达载体, 经农杆菌介导转化水稻, 获得超表达OsPR10A的纯合株系。田间表型观察表明, 转基因株系株高变矮、穗长变短、结实率降低。用20% PEG6000在水稻种子萌发过程中进行干旱处理, 结果显示, OsPR10A超表达株系的根长和芽长均显著高于野生型, 证明超表达OsPR10A可增强水稻萌发期耐旱性。该研究有助于增进人们对水稻OsPR10A功能的了解。

关键词: 水稻, 干旱胁迫, 病程相关蛋白, 免疫印迹, 基于抗体的蛋白质组学

Abstract: In this study, we surveyed the expression patterns of rice OsPR10A protein by using western blot analysis for protein samples extracted from different tissues at different developmental stages and a variety of abiotic stresses. The expression of OsPR10A was upregulated significantly under drought, salt stress and hormone treatments such as methyl jasmonate and abscisic acid, which suggests that OsPR10A protein plays a role in drought and salt stress response. To test this hypothesis, we constructed a vector for overexpression of OsPR10A protein and obtained homozygous transgenic lines via Agrobacterium-mediated rice transformation. Phenotypic investigation in field experiments revealed shorter plant height and spike length as well as lower seed setting rate in transgenic lines than controls. At seed germination stage, drought stress treatment was applied by using 20% PEG6000. Root and shoot length was significantly longer in OsPR10A overexpression lines than the wild-type control, which supports that overexpression of OsPR10A protein enhanced the drought tolerance of rice during germination. The study increases our understanding of the function of rice OsPR10A protein.

Key words: rice, drought stress, pathogenesis-related protein, western blot, antibody-based proteomics

张彤,郭亚璐,陈悦,马金姣,兰金苹,燕高伟,刘玉晴,徐珊,李莉云,刘国振,窦世娟. 水稻OsPR10A的表达特征及其在干旱胁迫应答过程中的功能. 植物学报, 2019, 54(6): 711-722.

Tong Zhang,Yalu Guo,Yue Chen,Jinjiao Ma,Jinping Lan,Gaowei Yan,Yuqing Liu,Shan Xu,Liyun Li,Guozhen Liu,Shijuan Dou. Expression Characterization of Rice OsPR10A and Its Function in Response to Drought Stress. Chinese Bulletin of Botany, 2019, 54(6): 711-722.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB19042

https://www.chinbullbotany.com/CN/Y2019/V54/I6/711

图/表 6

图1 OsPR10A在水稻生长发育过程中的表达 (A) OsPR10A在水稻不同生长时期和组织部位的表达(Sd: 苗期; Ti: 分蘖期; Bt: 孕穗期; Fw: 开花期); (B) OsPR10A在萌发期种子中的表达; (C) OsPR10A在开花期组织中的表达(Sp: 穗; Ss: 穗轴; An: 花药; Husk: 颖壳; Low: 叶下部; Mid: 叶中部; Up: 叶上部); (D) OsPR10A在幼苗和旗叶中的表达

Figure 1 Expression of OsPR10A during rice growth and development (A) Expression of OsPR10A in different growth stages and tissue parts of rice (Sd: Seedling stage; Ti: Tillering stage; Bt: Booting stage; Fw: Flowering stage); (B) Expression of OsPR10A in seeds during germination; (C) Expression of OsPR10A in tissues during flowering stage (Sp: Spike; Ss: Spike-stalk; An: Anther; Husk: Husk; Low: Lower part of leaf; Mid: Middle part of leaf; Up: Upper part of leaf); (D) Expression of OsPR10A in seedling and flag leaf

表1 OsPR10A基因的转录分析

Table 1 Transcriptome analysis of the OsPR10A gene

Library description	FPKM
Shoots	50.1661
Leaves-20 day	144.7290
Pre-emergence inflorescence	4.1989
Post-emergence inflorescence	88.9886
Anther	0.5100
Pistil	2.3483
Seed-5 DAP	77.4876
Seed-10 DAP	0.0000
Embryo-25 DAP	1.5052
Endosperm-25 DAP	2.3371

图2 水稻OsPR10A在非生物胁迫下的动态表达 (A) OsPR10A在干旱胁迫下的表达; (B) OsPR10A在盐胁迫下的表达; (C) OsPR10A在外源施加茉莉酸甲酯后的表达; (D) OsPR10A在外源施加脱落酸后的表达。MeJA: 茉莉酸甲酯; ABA: 脱落酸

Figure 2 Dynamic expression of rice OsPR10A under abiotic stress (A) Expression of OsPR10A under drought stress; (B) Expression of OsPR10A under salt stress; (C) Expression of OsPR10A modulated by exogenous MeJA; (D) Expression of OsPR10A modulated by exogenous ABA. MeJA: Methyl jasmonate; ABA: Abscisic acid

图3 超表达OsPR10A转基因水稻的鉴定 A721、A726、A728和A730分别代表不同转基因株系; 泳道1-7或1-8分别代表同一转基因株系的不同植株; WT: 野生型

Figure 3 Identification of OsPR10A overexpression transgenic rice A721, A726, A728 and A730 represent different transgenic lines, respectively; Lane 1-7 or 1-8 represent different plants of the same transgenic line, respectively; WT: Wildtype

图4 超表达OsPR10A转基因水稻的表型和农艺性状 (A) 成熟期植株(Bar=10 cm); (B) 穗部(Bar=4 cm); (C)-(F) 分别表示株高、穗长、结实率和分蘖数。WT: 野生型。* P<0.05; ** P<0.01

Figure 4 Phenotype of OsPR10A overexpression transgenic rice (A) Mature plant (Bar=10 cm); (B) Spike (Bar=4 cm); (C)-(F) Represent plant height, spike length, seed setting rate and tiller number, respectively. WT: Wildtype. * P<0.05; ** P<0.01

图5 超表达OsPR10A增强水稻种子萌发期的耐旱性 (A) 干旱处理(20% PEG6000) 7天萌发后的水稻种子(Bar=1 cm); (B) 根长; (C) 芽长。WT: 野生型。* P<0.05; ** P<0.01

Figure 5 Overexpression of OsPR10A enhanced the drought tolerance at rice seed germination stage (A) The germinated rice seeds at 7 days treated with 20% PEG6000 (Bar=1 cm); (B) Root length; (C) Shoot length. WT: Wildtype. * P<0.05; ** P<0.01

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水稻OsPR10A的表达特征及其在干旱胁迫应答过程中的功能

Expression Characterization of Rice OsPR10A and Its Function in Response to Drought Stress

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