| 王艳丽,谢天,张春丽,李娟娟,刘溢健,李红兵,殷俐娜,王仕稳,邓西平,可庆波.马铃薯气孔密度表皮模式因子StSTOMAGEN的克隆和功能分析[J].干旱地区农业研究,2020,38(4):235~244 |
| 马铃薯气孔密度表皮模式因子StSTOMAGEN的克隆和功能分析 |
| Cloning and functional analysis of potato stomatal density epidermal patterning factor StSTOMAGEN |
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| DOI：10.7606/j.issn.1000-7601.2020.04.30 |
| 中文关键词: 气孔密度 StSTOMAGEN 抗旱性 马铃薯 |
| 英文关键词:stomatal density StSTOMAGEN drought tolerance potato |
| 基金项目:中国科学院“黄土高原土壤侵蚀与旱地农业国家重点实验室”开放基金（A314021402-2013）；中国科学院“西部青年学者”B类项目（XAB2017B02） |
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| 中文摘要: |
| 为了研究马铃薯EPFL9/StSTOMAGEN基因在气孔发育过程中的功能，从马铃薯品种大西洋中克隆得到气孔密度表皮模式因子StSTOMAGEN，将其通过农杆菌介导的遗传转化方法，在拟南芥中异位表达，对该基因的表达特性及功能进行分析。结果显示：StSTOMAGEN主要定位于细胞间隙和细胞核，并且它在顶端未展开叶中表达最多，表达量在1 d中呈现周期性变化，长日照条件下（16 h/8 h，光/暗），在授时因子为20 h时表达量最高；聚乙二醇（PEG）模拟干旱胁迫处理4 h后其表达量达到最高，而脱落酸（ABA）和NaCl胁迫处理后，其表达水平呈现出持续降低的趋势。功能鉴定表明，过表达StSTOMAGEN基因的拟南芥（ST）相对于野生型（WT）气孔密度增大63%~83%，光合速率增大36%~42%。此外，ST植株离体叶片的水分散失速率增大24%~55%，离体叶片H2O2含量也显著升高，约为野生型的50%~100%。干旱胁迫下，ST株系的光合速率、Fv/Fm和瞬时水分利用效率都显著低于野生型，其中光合速率降低55%~66%，Fv/Fm降低22%~50%，瞬时水分利用效率降低11%~12%。综上结果可知，表皮模式因子StSTOMAGEN通过正调控气孔密度而降低植物的抗旱性，该结果为后期通过基因编辑技术降低StSTOMAGEN基因的表达，培育抗旱节水型马铃薯新品种奠定理论基础。 |
| 英文摘要: |
| Secretion of small molecules in the epidermal patterning factor\|like family (EPFL) play an important role in stomatal development. To study the function of potato EPFL9/StSTOMAGEN gene during stomatal development, we cloned and functionally characterized the stomatal density regulator StSTOMAGEN from potato (Atlantic) and transformed it into Arabidopsis by Agrobacterium\|mediated genetic transformation. We found StSTOMAGEN was mainly inter cellular space and cell nucleus localized and had the highest expression level in the apical unexpanded leaves. Interestingly, StSTOMAGEN transcript was rhythmically expressed with a peak around Zeitgeber time (ZT) 20 under long\|day (16 h/8 h, light/dark) conditions. Expression of StSTOMAGEN was markedly down\|regulated in response to abscisic acid (ABA) and sodium chloride (NaCl) treatments; however, upon the application of polyethylene glycol (PEG), the expression of StSTOMAGEN peaked at 4 h and then decreased gradually. Functional identification showed that the stomatal density of the StSTOMAGEN overexpression (ST) lines was significantly increased by 63%~83%, and led to elevated photosynthetic rateby 36%~42%. Moreover, ST lines showed more water loss (24%~55%) and hydrogen peroxide (H2O2) accumulation (50%~100%) in detached leaves compared with wild\|type (WT) plants. Under drought stress, ST lines maintained lower photosynthetic rates (55%~66%), photosystem Ⅱ efficiency (22%~50%), and instantaneous water use efficiency (111%~12%) than WT plants did. Taken together, the results showed that the epidermal patterning factor StSTOMAGEN reduced drought tolerance of plants by positively regulating stomatal density. This result laid a theoretical foundation for reducing the expression of StSTOMAGEN gene and cultivating drought\|resistant and water\|saving potato strains by gene editing technology in the later period. |
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