水稻AGPase小亚基基因的克隆测序及种质间遗传分化的研究

淀粉是构成和影响水稻产量和品质的最主要因素;ADP-葡萄糖焦磷酸化酶(AGPase)基因是影响淀粉合成的关键酶基因之一。本研究用编码AGPase小亚基基因(AGPsma)序列的特异引物(序列为F:5’-TACGCTATGCTCTTGAAAC-3’;R:5’-TATCTTCCCAGTAACCATCA-3’)对普通野生稻(元江)、粳稻(榆密15)、籼稻(93-11)及籼粳交品系(南34)进行PCR扩增、克隆、测序和序列对比。又用该引物对来源于13个国家的15份AA基因组和5份CCDD基因组的共20份野生稻及205份包括40份籼稻和165份粳稻的云南地方品种和改良品种(系)进行PCR检测。在以上4个不同类型水稻材料该引物上扩增出184bp和215bp的2个片段均为AGPase小亚基基因的序列。在该序列的109bp处,元江普通野生稻和榆密15为T,而93-11和南34为C;籼粳交品系南34在121~151bp区间缺失31个碱基。所有的225份供试材料可分为3类。第一类具有215bp大小的条带,包括了全部的野生稻材料和196份籼粳材料。其中籼稻数占籼稻样本总数的95%,籼稻地方品种占籼稻地方品种总数的75%;粳稻数占粳稻样本总数的83.6%,粳稻地方品种占粳稻地方品种总数的60%。第二类具有184bp的条带,包括28个籼粳材料。第三类同时具有215bp和184bp的片段,此类只有1个父母本具不同片段的滇型杂交粳稻品种。结果表明AGPsma序列在野生稻中可能尚未出现分化,但在籼稻、粳稻和地方老品种及改良品种中都出现了遗传分化。

Cloning and Sequencing of Rice ADP-Glucose Pyrophosphorylase Small Subunit Gene and Study on Genetic Transformation among Germplasm

The quality and yi eld of rice are characterized by starch. ADP-glucose pyrophosphorylase small subunit gene is one of the key enzyme genes of starch synthesis. In this study, PCR amplification, cloning, sequencing and sequence alignment of Common wild rice (from Yuanjiang), japonica (Yumi15), indica (93-11) and indica-japonica lines (Nan34) were analyzed by specific primer (sequences of F: 5’-TACGCTATGCTCTTGAAAC- 3’; R: 5’-TATCTTCCCAGTAACCATCA-3’) to ADP-glucose pyrophosphorylase small subunit gene (AGPsma). Besides, 20 accessions of wild rice of 15 AA and 5 CCDD genomes from 13 countries, and 205 rice cultivars which covers 40 Indica and 165 Japonica including Yunnan landraces and improved lines were PCR detected by the primer. The fragments of 184 bp and 215 bp in four different rice with the primer were sequences of AGPsma. The similarity between common wild rice (from Yuanjiang) and Yumi15, between common wild rice and 93-11, and between common wild rice and Nan34 was 100%, 99.54%, and 85.19%, respectively. On 109 bp of the sequence, common wild rice from Yuanjiang and Yumi15 were T, but 93-11 and Nan34 were C. In the range of 121~151 bp, indica-japonica lines Nan34 deleted 31 bases. The total rice accessions were divided intothree classes. The first class only has 215 bp fragment, including all the wild rice and 196 indica and Japonica materials. The number of indica accounted for 95% of the total indica samples, and indica landraces accounted for 75% of the total indica landraces; The number of Japonica accounted for 83.6% of the total Japonica samples, and Japonica landraces accounted for 60% of the total Japonica landraces; The second class only has 184 bp fragment, including 28 and Japonica materials. The third class have 215 bp and 184 bp fragments, including only one japonica hybrid material, the fragments of which parents were diffenent. The results indicated that genetic differential of the sequence of AGPsma has not yet appeared in the wild rice, but it appeared between indica and Japonica and between Yunnan landraces and improved lines.