PDF
Full
Outline
Cassava is an important cash crop in Southern China, which is enriched with starch in the tuber roots. The analysis of the regulation mechanism of starch synthesis in cassava root would contribute to the improvement of yield and high starch molecular in cassava. AGPase is composed of large subunit and small subunit, and it catalyzes G-1-P and ATP to form ADPG and PPi, in which ADPG is the substrate of starch biosynthesis. Therefore, AGPase is a rate-limiting enzyme in plant starch synthesis, and improving the activity of AGPase is beneficial for the accumulation of crop starch and the improvement of yield. The small subunit encoded by MeAGPS1a is the catalytic center of AGPase in cassava. Previous studies have shown that the growth response factor MeSAUR1 as a transcription factor positively regulates the expression of MeAGPS1a gene, and yeast two-hybrid screening of a cassava cDNA library revealed that the calmodulin-like (CML) member MeCML24 is a candidate interacting protein of MeSAUR1. To determine the interaction between MeCML24 and MeSAUR1, and MeCML24 was cloned from the genome of SC8 cassava variety in this study. The length of CDS region of MeCML24 was 492 bp, encoding 163 amino acids. The physicochemical properties and secondary structure analysis of MeCML24 showed that the value of theoretical pI was 4.38, which belonging to hydrophilic protein with α-helix accounting for 52.76%, random coil accounting for 30.06%, and beta turn accounting for 11.04%. Yeast two-hybrid vector BD-MeCML24 was constructed, and the self-activation experiment showed that MeCML24 had no self-activation. Yeast two-hybrid point-to-point experiment revealed that yeast strains co-transfected with AD-MeSAUR1 and BD-MeCML24 plasmids turned blue on nutrient medium with SD/TLHA+x-α-gal, indicating that MeCML24 interacted with MeSAUR1. The fusion protein MeSAUR1-nEYFP and MeCML24-cEYFP were co-expressed in tobacco leaves by bimolecular fluorescence complementation (BiFC) experiment, and the fluorescence signal of yellow fluorescent protein EYFP was detected under laser confocal microscope, which further proved the interaction between MeCML24 and MeSAUR1. Finally, the interaction between MeCML24 and MeSAUR1 negatively regulated the expression of MeAGPS1a by double luciferase experiment. The study revealed the mechanism of MeSAUR1 and MeCML24 synergistically regulating MeAGPS1a gene expression in cassava, and found that the calmodulin-like protein MeCML24 regulating the expression of MeAGPS1a, a key gene for starch synthesis in cassava root. This study would provide a theoretical basis for cultivating excellent cassava varieties by molecular biology technology.
PDF
49
16
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
关闭全屏
No. 86 Xueyuan South Road, Haidian District, Beijing
010-62199257
qkjq@cast.org.cn
Copyright © 2025 China Association for Science and Technology. All rights reserved. For all open access content, the relevant licensing terms apply.
Sponsored by the Office of the Leading Group for Cybersecurity and Informatization of CAST, and supported by Science and Technology Review Publishing House
