crispr - cas9介导的ZmPL1基因编辑提高玉米对干旱胁迫的耐受性

Chunlai Wang,Yangyang Zhou,Yimeng Wang,Peng Jiao,Siyan Liu,Shuyang Guan,Yiyong Ma
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摘要

玉米(Zea mays L.)是世界上广泛种植的粮食作物。干旱胁迫严重影响植物的生长发育过程,对玉米产量造成严重损害。课题组前期对抗旱玉米自交系H8186进行转录组测序分析,筛选出一个表达显著下调的基因Phylloplanin-like (ZmPL1)。分析了ZmPL1基因在不同非生物胁迫下的表达谱,结果表明该基因受干旱胁迫影响较大。亚细胞定位分析表明,该蛋白定位于细胞膜上。为了验证ZmPL1在干旱胁迫中的作用,我们在酵母中过表达ZmPL1,发现表达ZmPL1可以显著提高酵母的干旱敏感性。接下来,利用农杆菌介导法侵染玉米愈伤组织,获得ZmPL1转基因植株。在干旱胁迫下,与过表达品系相比,基因编辑品系的发芽率和幼苗成活率更高,MDA、相对电导率和ROS积累更低,抗氧化酶活性更高,胁迫相关基因和ROS清除相关基因的表达水平显著增加。在干旱胁迫下,各株系外源施用ABA可减轻干旱胁迫对ZmPL过表达植株造成的伤害。综上所述,ZmPL1负调控玉米抗旱性。
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CRISPR-Cas9-mediated editing of ZmPL1 gene improves tolerance to drought stress in maize.
Maize (Zea mays L.) is a widely grown food crop around the world. Drought stress seriously affects the growth and development process of plants and causes serious damage to maize yield. In the early stage, our research group conducted transcriptome sequencing analysis on the drought-resistant maize inbred line H8186 and screened out a gene with significantly down-regulated expression, Phylloplanin-like (ZmPL1). The ZmPL1 gee expression pattern was analyzed under various abiotic stresses, and the results showed that this gene was greatly affected by drought stress. Subcellular localization analysis showed that the protein was localized on the cell membrane. In order to verify the role of ZmPL1 in drought stress, we overexpressed ZmPL1 in yeast and found that the expression of ZmPL1 could significantly increase the drought sensitivity of yeast. Next, ZmPL1 transgenic plants were obtained by infecting maize callus using Agrobacterium-mediated method. Under drought stress, compared with overexpression lines, gene-edited lines had higher germination rate and seedling survival rate, lower accumulation of MDA, relative conductivity and ROS, higher antioxidant enzyme activity, and the expression levels of stress-related genes and ROS scavenging-related genes were significantly increased. Exogenous application of ABA to each lines under drought stress attenuated the damage caused by drought stress on ZmPL overexpressing plants. In summary, ZmPL1 negatively regulates drought tolerance in maize.
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