New Phytol：中農丨PTP3ases調節棉花耐鹽新機制（附NMT實驗體系）

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期刊：New Phytologist

主題：PTP3ases調節棉花耐鹽新機制

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

影響因子：7.299

檢測指標：Ca2+、Na+、K+流速

Ca2+流實驗方法：

7d擬南芥，100 mM NaCl瞬時鹽脅迫處理，檢測距根尖1000微米的點

Ca2+流實驗測試液成份：
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+流實驗方法：

· 5d擬南芥，0、100 mM NaCl處理24h，檢測距根尖200微米的點
· 6d擬南芥，50 uM LaCl3+100 mM NaCl處理24h，檢測距根尖200微米的點

K+、Na+流實驗測試液成份：
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：中國農業大學李芳軍、穆春、得州農工大學單麗波

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（谷歌機翻）

鹽度是限制全球作物生產的主要因素之一。盡管具有適度的耐鹽性，但棉花（Gossypium spp。）遭受鹽度脅迫的嚴重產量損失，主要是由于在鹽堿地和干旱地區生長。

為了鑒定賦予棉花耐鹽性的遺傳決定因子，我們在病毒誘導的基因沉默（VIGS）載體中使用棉花cDNA文庫部署了功能基因組篩選。我們已經發現，編碼蛋白磷酸酶的GhDsPTP3a的沉默會增加棉花對鹽脅迫的耐受性。

酵母雙雜交篩選表明GhDsPTP3a與膜聯蛋白GhANN8b相互作用，后者在調節棉花對鹽脅迫的反應中起著積極作用。鹽脅迫誘導GhANN8b磷酸化，其隨后被GhDsPTP3a去磷酸化。GhDsPTP3a和GhANN8b的異位表達相反地調節植物鹽耐受性和鈣內流。此外，我們發現GhDsPTP3a或GhANN8b的沉默在調節GhSOS1轉錄水平中發揮相反的作用，并且GhANN8b的異位表達在鹽脅迫下提高擬南芥中的Na+流出。

我們的研究表明棉花磷酸酶GhDsPTP3a和膜聯蛋白GhANN8b在棉花鹽度反應中相互作用并反向調節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.

文章鏈接：

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