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作物学报 ›› 2020, Vol. 46 ›› Issue (6): 809-818.doi: 10.3724/SP.J.1006.2020.91067

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

小麦脱落酸受体基因TaPYR1介导植株抵御干旱逆境功能研究

韩乐,杜萍萍,肖凯()   

  1. 河北农业大学农学院, 河北保定071001
  • 收稿日期:2019-11-09 接受日期:2020-01-15 出版日期:2020-06-12 网络出版日期:2020-02-14
  • 通讯作者: 肖凯
  • 作者简介:E-mail:hanle95@163.com
  • 基金资助:
    国家自然科学基金项目(31872869);国家重点研发计划项目(2017YFD0300902)

Functional characteristics of TaPYR1, an abscisic acid receptor family gene in mediating wheat tolerance to drought stress

HAN Le,DU Ping-Ping,XIAO Kai()   

  1. College of Agronomy, Hebei Agricultural University, Baoding 071001, Hebei, China
  • Received:2019-11-09 Accepted:2020-01-15 Published:2020-06-12 Published online:2020-02-14
  • Contact: Kai XIAO
  • Supported by:
    Natural Science Foundation of China(31872869);National Key Research and Development Program of China(2017YFD0300902)

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摘要：

脱落酸(ABA)受体PYR/PYL/RCAR通过与渗透胁迫诱导的ABA结合, 参与植株体内ABA介导的信号转导过程, 在调控植株干旱逆境抵御过程中具有重要的生物学功能。本文研究了鉴定的1个干旱响应的PYR家族成员TaPYR1的分子特征、应答干旱表达模式及其介导植株抵御干旱逆境的功能。结果表明, TaPYR1与植物种属中部分PYR基因在氨基酸序列水平上高度同源, 编码蛋白含有PYR家族成员保守结构域, 翻译蛋白经内质网分选后定位于细胞质膜。TaPYR1基因在小麦根、叶中均呈明显的干旱诱导表达模式, 在干旱胁迫48 h表达量达到峰值。与野生型对照(WT)相比, 超表达TaPYR1烟草转化株系, 干旱处理下植株长势增强, 干鲜重增加。干旱处理下, 转化株系较对照的光合能力增强, 细胞保护酶活性提高, 渗透调节物质脯氨酸和可溶性糖含量增加。研究表明, TaPYR1通过在转录水平上应答干旱逆境, 改善干旱胁迫下的植株相关生理过程, 在增强植株抵御干旱逆境能力中发挥重要作用。

关键词: 小麦, ABA受体基因, 表达, 遗传转化, 功能鉴定

Abstract:

ABA receptors are involved in the mediation of ABA signaling transduction through interaction with abscisic acid (ABA) molecules induced by osmotic stresses and play critical roles in regulating the drought stress tolerance in plants. In this study, TaPYR1, an ABA family gene in wheat that was shown to be differentially expressed in our previous transcriptome analysis was used to analyze its molecular property, expression patterns under drought stress condition, and functions in mediating plant adaptation to drought stress. TaPYR1 shares high similarities to its plant counterparts at amino acid level. TaPYR1 protein contains the conserved domains specified by the plant PYR proteins and was targeted onto the plasma membrane after endoplasmic reticulum (ER) assortment. The expression of TaPYR1 was induced in both roots and leaves under drought, with the highest expression levels at 48 h of drought treatment. Transgene analysis on TaPYR1 was performed to assess the gene function in mediating plant drought tolerance. Compared with wild type (WT), the tobacco lines overexpressing TaPYR1 enhanced growth vigor and increased fresh and dry weight under drought stress. In addition, the transgenic lines with TaPYR1 overexpression also increased photosynthetic function, enhanced activities of cellular antioxidant enzymes, and elevated the contents of osmolytes (i.e., proline and soluble sugar) under drought condition. Our investigation suggests that TaPYR1 transcriptively responds to drought stress signaling and plays an important role in regulating plant drought adaptation by improving the associated physiological processes.

Key words: Triticum aestivum, ABA receptor gene, expression, genetic transformation, functional characterization

附表1

NtSOD、NtCAT和NtPOD家族基因特异性扩增引物"

目的基因
Gene		 登录号
Accession number		 上游引物
Forward primer (5'-3')		 下游引物
Reverse primer (5'-3')
NtSOD1 KJ874395 GTGGACATGTCGTGTCAAGG TTCTCACCAACTCCTGCACTT
NtFeSOD KF724056 CATCACAGAGCTTATGTCGACA CTAGAACTGACTGCTTCCCA
NtSOD2 EU123521 ATGTCACGGGACCACATTAC AACCCTTCCACCAGCATTTC
NtMnSOD2 AB093097 GACGGACCTTAGCAACAGGG ACCAATGGGTCCTGATTAGCAG
NtCAT NTU07627 GTCTCAGGCTGACAAGTCTT ACGGAAGACAGAGTAGCAGC
NtCAT1 EF532799 CAAGGATCTCTACGACTCGATT CTTGAGGGCAAATAATCCACCT
NtCAT1;1 NTU93244 TCCTGCTAATGCTCCAAAGTGT AATGCATATGTATTAGGAATGCTC
NtCAT1;2 HF564632 GGTATCGACTTGGACCAAACTA GGTCTCACATTAAGCCTAGAAG
NtCAT1;3 HF564631 TTGCAGCCGGTGGGAAGATT GGTCTCACATTAAGCCTAGAAG
NtCAT3 HF564633 GTCTTGGGCCAAACTATCTGCA TCAGCTTCACATTGTGGGCC
NtPOD1;1 L02124 GGAATTTGTCCTCAAGGTGGAA CTTATTGGAATTGCCATTTCAGC
NtPOD1;2 AB044154 CTGACATGGTCTGTGCCTAC TCAGTTGATAGCAGAGCAAACTT
NtPOD1;3 AB044153 AAGATCTTGTCGCTCTTACTGG AATTGGATTTTCCAGCTTGCG
NtPOD1;4 D11396 TGCTGGTAGTCAAAGTCAGTTTT CCCATGTTGAACACGTTCTTACC
NtPOD1;5 AB178953 AACAGCAACAACGTTAACCCAGC TTAATTTTGGACCACATTCAGGA
NtPOD1;6 AB027753 TCAACTCCACTGGTGGCCCT AATTCGATTTTGCAGCTTGCGC
NtPOD1;7 AB027752 GCCCAAGAAGTTCAGGCTCA ATACAAATACAGTCCTTTACTCG
NtPOD2;1 AB178954 AGGGGAAAAGACCTCACCAC AGTTTCCCATCTTGATCATAGCA
NtPOD4 AY032675 AGACTCAAAGATAGCAAACCTCA CTTCCTGATGTCACCCTTGA
NtPOD9 AY032674 CACCACCTTCATTCAACGCTA ACATCTCAGACAAAACACTTGTC

图1

TaPYR1蛋白的分子特征 A: TaPYR1蛋白结构预测; B: TaPYR1蛋白多重序列比对; C: TaPYR1氨基酸水平系统进化分析。"

图2

根叶中TaPYR1应答干旱胁迫逆境表达模式 0 h, 处理前对照; 6、12、24和48 h, 干旱处理时间点; R24 h和R48 h, 恢复处理时间点。"

图3

干旱胁迫下转化株系的植株生长特征 A: 干旱胁迫下TaPYR1转化株系植株生长情况; B: 干旱胁迫下TaPYR1转化株系干质量。WT, 野生型; Sen 1和Sen 2, 正义表达TaPYR1株系; Anti 1和Anti 2, 反义表达TaPYR1株系。误差线表示标准差, 符号*表示相对于WT的差异显著(P < 0.05)。"

图4

干旱胁迫下转化株系的光合特性 A: 光合速率(Pn); B: 胞间CO2浓度(Ci); C: 叶绿体光化学淬灭系数 (NPQ); D: 光系统II光化学效率(ФPSII)。误差线表示标准差, 符号*表示相对于WT的差异显著(P < 0.05)。"

图5

干旱胁迫下转化株系的气孔关闭及叶片持水特征 A: 气孔关闭特征; B: 叶片持水力。"

图6

干旱胁迫下转化株系的细胞保护酶活性及渗透物质含量 A: SOD活性; B: CAT活性; C: POD活性; D: MDA含量; E: 脯氨酸含量; F: 可溶性糖含量。误差线表示标准差, 符号*表示相对于WT的差异显著(P < 0.05)。"

图7

干旱胁迫下转化株系的保护酶基因表达模式 A: SOD家族基因; B: CAT家族基因C: POD家族基因。误差线表示标准差, 符号*表示相对于WT的差异显著(P < 0.05)。"

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