New Phytol:中农丨PTP3ases调节棉花耐盐新机制(附NMT实验体系)

来源: 旭月(北京)科技有限公司   2019-7-5   访问量:443评论(0)

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期刊:New Phytologist

主题:PTP3ases调节棉花耐盐新机制

标题:Phosphatase GhDsPTP3a interacts with annexin protein GhANN8b to reversely regulate salt tolerance in cotton (Gossypium spp.)

影响因子:7.299

检测指标:Ca2+、Na+、K+流速

Ca2+流实验方法:

7d拟南芥,100 mM NaCl瞬时盐胁迫处理,检测距根尖1000微米的点

Ca2+流实验测试液成份:
0.1 mM KCl, 0.1 mM CaCl2, 0.1 mM MgCl2, 0.5 mM NaCl, 0.3 mM MES, 0.2 mM Na2SO4, 0.1 % sucrose, pH 6.0

K+、Na+流实验方法:

· 5d拟南芥,0、100 mM NaCl处理24h,检测距根尖200微米的点
· 6d拟南芥,50 uM LaCl3+100 mM NaCl处理24h,检测距根尖200微米的点

K+、Na+流实验测试液成份:
0.5 mM KCl, 0.1 mM CaCl2, 0.1 mM MgCl2, 0.1 mM NaCl,
0.3 mM MES, pH 6.0

作者:中国农业大学李芳军、穆春得州农工大学单丽波


英文摘要

Salinity is among the major factorslimiting crop production worldwide. Despite having moderate salt‐tolerance,cotton (Gossypium spp.) suffers severe yield losses to salinity stresses,largely due to being grown on saline‐alkali and drylands.

To identify genetic determinants conferringsalinity tolerance in cotton, we deployed a functional genomic screen using acotton cDNA library in a virus‐induced gene silencing (VIGS) vector. We have revealed thatsilencing of GhDsPTP3a, which encodes a protein phosphatase, increases cottontolerance to salt stress.

Yeast two‐hybrid screensindicated that GhDsPTP3a interacts with GhANN8b, an annexin protein, whichplays a positive role in regulating cotton response to salinity stress. Saltstress induces GhANN8b phosphorylation, which is subsequently dephosphorylated byGhDsPTP3a. Ectopic expression of GhDsPTP3a and GhANN8b oppositely regulatesplant salt tolerance and calcium influx. In addition, we have revealed thatsilencing of GhDsPTP3a or GhANN8b exerts opposing roles in regulating GhSOS1transcript levels, and ectopic expression of GhANN8b elevates Na+ efflux inArabidopsis under salinity stress.

Our study demonstrates that a cottonphosphatase GhDsPTP3a and an annexin protein GhANN8b interact and reverselymodulate Ca2+ and Na+ fluxes in cotton salinity responses.

中文摘要(谷歌机翻)

盐度是限制全球作物生产的主要因素之一。尽管具有适度的耐盐性,但棉花(Gossypium spp。)遭受盐度胁迫的严重产量损失,主要是由于在盐碱地和干旱地区生长。

为了鉴定赋予棉花耐盐性的遗传决定因子,我们在病毒诱导的基因沉默(VIGS)载体中使用棉花cDNA文库部署了功能基因组筛选。我们已经发现,编码蛋白磷酸酶的GhDsPTP3a的沉默会增加棉花对盐胁迫的耐受性。

酵母双杂交筛选表明GhDsPTP3a与膜联蛋白GhANN8b相互作用,后者在调节棉花对盐胁迫的反应中起着积极作用。盐胁迫诱导GhANN8b磷酸化,其随后被GhDsPTP3a去磷酸化。GhDsPTP3a和GhANN8b的异位表达相反地调节植物盐耐受性和钙内流。此外,我们发现GhDsPTP3a或GhANN8b的沉默在调节GhSOS1转录水平中发挥相反的作用,并且GhANN8b的异位表达在盐胁迫下提高拟南芥中的Na+流出。

我们的研究表明棉花磷酸酶GhDsPTP3a和膜联蛋白GhANN8b在棉花盐度反应中相互作用并反向调节Ca2+和Na+通量。

(b) Ectopic-expression of GhDsPTP3a or GhANN8b alters extracellular Na+ efflux upon salt stress in live roots via NMT assay. (c) LaCl3 blocks GhANN8b-induced Na+ efflux upon salt stress in live roots via NMT assay.

文章链接:

https://nph.onlinelibrary.wiley.com/doi/pdf/10.1111/nph.15850#accessDenialLayout

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