摘要
【目的】盐害作为影响植物生长发育的非生物胁迫因子,严重威胁林木生长。在受到盐胁迫时,植物内源活性氧(ROS)水平增加,造成氧化胁迫,影响植株正常生长发育。因此,可通过增强过氧化物酶PRX家族成员表达水平,改变ROS水平,以增强杨树Populus耐盐能力,揭示PRX成员参与调控杨树盐胁迫响应的机制。【方法】以银腺杨‘84K’Populus alba×P. glandulosa ‘84K’为材料,生物信息学分析选取PRX家族成员PagPRX19进行克隆并构建过表达载体,农杆菌Agrobacterium tumefaciens介导叶盘转化法获得过表达植株。以银腺杨‘84K’ PagPRX19过表达植株生长45 d的组培苗和生长2个月的土培苗为实验材料,进行盐胁迫处理,以非转基因植株为对照。观察植株表型,检测脯氨酸、丙二醛、电解质渗透率等生理指标并进行分析。【结果】(1)克隆了PagPRX19基因,构建过表达载体,获得转基因阳性植株。经分子鉴定选取2个过表达株系OE#1和OE#2为实验材料做后续分析。(2)与对照相比,过表达植株株高下降,地径增加。(3)盐胁迫处理下,过表达植株相较于对照表现为叶片皱缩以及植株生长受到抑制程度低,组培苗的盐胁迫处理表现为相似结果。(4)转基因植株的ROS水平降低,而且在盐胁迫下过表达植株叶片和根的ROS仍保持较对照低的水平。盐胁迫下过表达植株较对照脯氨酸增加,叶片持水能力增强,丙二醛和电解质渗透率降低。从生理方面显示转基因植株具有较高的耐盐能力。【结论】过表达PagPRX19可降低盐胁迫下杨树转基因植株的ROS水平,缓解氧化胁迫,增强了植株耐盐性。图11参23。
[Objective] Salt damage, as an abiotic stress factor affecting plant growth and development,seriously threatens tree growth. Under salt stress, the level of endogenous reactive oxygen species(ROS) in plants increases, resulting in oxidative stress and affecting the normal growth and development of plants. By enhancing the expression level of PRX family members and changing ROS levels, the salt tolerance ability of poplar can be enhanced, and the mechanism of PRX members involved in regulating salt stress response can be revealed. [Method] Poplar ‘84K’(Populus alba × P. glandulosa ‘84K’) was used as material, and PagPRX19,a member of PRX family, was selected for cloning and construction of overexpression vector by bioinformatics analysis. Overexpressed plants were obtained by Agrobacterium tumefaciens mediated leaf disk transformation. The tissue culture seedlings of non-transgenetic plants(the control) and PagPRX19 overexpressed plants growing for 45 days and the soil culture seedlings growing for 2 months were used as experimental materials for salt stress treatment. The plant phenotype was observed and the physiological indexes such as proline,malondialdehyde and electrolyte permeability were detected and analyzed. [Result](1) PagPRX19 gene was cloned, overexpression vector was constructed, and transgenic positive plants were obtained. After molecular identification, 2 overexpressed lines OE#1 and OE#2 were used for further analysis.(2) Preliminary phenotypic observation showed that PagPRX19 overexpressed plants decreased in height and increased in ground diameter,compared with the control.(3) Compared with the control, the overexpressed plants showed lower leaf shrinkage and plant growth inhibition under salt stress. Tissue culture seedlings showed similar results under salt stress.(4) ROS level of transgenic plants decreased, and ROS level of leaves and roots of overexpressed plants remained lower than that of the control under salt stress. The content of proline in overexpressed plants increased, th
作者
黄清晨
赖建新
黄李超
卢孟柱
HUANG Qingchen;LAI Jianxin;HUANG Lichao;LU Mengzhu(College of Forestry and Biotechnology,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;State Key Laboratory of Subtropical Silviculture,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China)
出处
《浙江农林大学学报》
CAS
CSCD
北大核心
2022年第6期1163-1172,共10页
Journal of Zhejiang A&F University
基金
国家重点研发计划项目(2019YFE0119100)。
