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MeLAZY1c基因调控木薯株型的初步研究

望嘉翔1，郁雪婷1，李梦桃1，麦伟涛1，陈新2,3,*，王文泉1,*   

  1. 1 海南大学热带农林学院 / 三亚南繁研究院, 海南三亚 572000；2 中国热带农业科学院热带生物技术研究所 / 海南热带农业资源研究院海南省热带农业生物资源保护与利用重点实验室, 海南海口 571101；3 中国热带农业科学院三亚研究院, 海南三亚 572000
  • 收稿日期:2023-09-12 修回日期:2024-01-12 接受日期:2024-01-12 网络出版日期:2024-02-09
  • 基金资助:
    本研究由国家自然科学基金NSFC-CG联合基金重点项目(3181101517)，国家重点研发计划项目(2018YFD1000500)和海南省高校研究生创新科研课题(Qhys2022-82)资助。

Preliminary study on the regulation of cassava plant type by MeLAZY1c gene

WANG Jia-Xiang1, YU Xue-Ting1, LI Meng-Tao1, MAI Wei-Tao1, CHEN Xin2,3,*,WANG Wen-Quan1,*   

  1. 1 College of Tropical Agriculture and Forestry / Sanya Nanfan Research Institute, Hainan University, Sanya 572000, Hainan, China; 2 Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences / Hainan Key Laboratory of Conservation and Utilization of Tropical Agricultural Biological Resources, Hainan Institute of Tropical Agricultural Resources, Haikou 571101, Hainan, China; 3 Sanya Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572000, Hainan, China
  • Received:2023-09-12 Revised:2024-01-12 Accepted:2024-01-12 Published online:2024-02-09
  • Supported by:
    This study was supported by the National Natural Science Foundation of China NSFC-CG Joint Fund Key Project (3181101517), the National Key Research and Development Program of China (2018YFD1000500) and the Innovation Research Project for Graduate Students in Hainan Province (Qhys2022-82).

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摘要：

IGT基因家族参与作物株型的调控，LAZY属于IGT的亚家族。以拟南芥6个LAZY成员氨基酸序列为“种子”在木薯基因组中进行比对，在木薯中共鉴定到8个LAZY基因，其中MeLAZY1c与调控分枝角度的AtLAZY1高度同源。基于此，本研究以MeLAZY1c为研究对象，利用qRT-PCR分析发现MeLAZY1c在茎中转录水平最高，GUS染色显示pMeLAZY1c在维管束中染色较深。在MeLAZY1c启动子中发现8个光响应/调节元件，随后发现黑暗能显著抑制其表达水平。同时对MeLAZY1c进行基因编辑，获得纯合编辑株系19个，炼苗移栽后观测表型，发现melazy1c突变体植株与SC8野生型相比，其主茎呈现匍匐生长，并且弯曲部位茎外皮细胞形态扭曲变形且大小不一致，近地侧1 mm处细胞数目约是远地侧细胞数量的1.5倍，表明MeLAZY1c在木薯直立/匍匐生长建成方面发挥着重要作用。

关键词: 木薯, 株型调控, LAZY, 光响应, 匍匐, 基因编辑

Abstract:

The IGT gene family is involved in the regulation of crop plant type, and LAZY belongs to the IGT subfamily. By comparing the amino acid sequences of six Arabidopsis LAZY members as ‘seeds’ in the cassava genome, a total of 8 LAZY genes were identified in cassava, among which MeLAZY1c is highly homologous to AtLAZY1, which regulates branching angles. Based on this, MeLAZY1c had the highest transcription level in the stem by qRT-PCR and pMeLAZY1c was deeply stained in the vascular bundle by GUS staining, using MeLAZY1c as the experimental materials in this study. Eight photoresponsive/regulatory elements were found in the MeLAZY1c promoter, and we subsequently found that darkness significantly inhibited the relative expression level of MeLAZY1c. At the same time, gene editing of MeLAZY1c was performed and 19 homozygous edited lineages. The phenotype of MeLAZY1c mutants was observed after seedling transplantation, compared to the SC8 wild-type, the main stems of MeLAZY1c mutants had crawing growth, and the stem skin cells at the curved part were distorted and deformed with different sizes. The number of cells at 1 mm near the ground was about 1.5 times that of cells at the far ground. In conclusion, these results indicates that MeLAZY1c plays an important role in the establishment of upright/creeping growth of cassava.

Key words: cassava, plant type regulation, LAZY, light response, crawling, gene editing

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