四倍体水稻蛋白质含量和谷蛋白基因表达研究

doi:10.7668/hbnxb.20192375

摘要/Abstract

摘要： 为了探讨高产四倍体水稻的不同遗传组成与品质的关系，以及蛋白质含量变化的原因，以国家优质品种美香占2号（MXZ2）、象牙香占（XYXZ）、矮脚南特二倍体（AJNT-2x）、高产同源四倍体水稻矮脚南特（AJNT-4x）以及高产新型四倍体水稻华多1号（H1）为试验材料，利用BCA法测定胚乳蛋白质含量、近红外谷物分析仪进行品质分析、全自动氨基酸分析仪测定氨基酸含量、qRT-PCR分析谷蛋白基因的表达差异。结果表明： AJNT-4x糙米、精米的蛋白含量分别为182.70，115.70 mg/g，高于国家优质品种MXZ2、XYXZ以及AJNT-2x，其营养品质大大提高；AJNT-4x的胶稠度为44.70 mm，低于MXZ2、XYXZ及AJNT-2x，且差异显著，其食用品质明显下降；而H1精米的蛋白含量最低，只有71.30 mg/g，但其胶稠度为112.70 mm，介于国家优质品种MXZ2与XYXZ之间。花后30 d AJNT-4x的17种氨基酸总量与人体必需氨基酸总量高于AJNT-2x与H1，H1的最低；H1的17种氨基酸总含量在花后15 d达到高峰，而AJNT-4x和AJNT-2x在花后20 d达到高峰，氨基酸含量积累过早停止导致H1在成熟期的总氨基酸含量下降，甚至低于AJNT-2x。在整个胚乳发育过程中，AJNT-4x、H1和AJNT-2x的4种贮藏蛋白积累趋势基本一致，但快速积累时间存在差异；AJNT-4x的谷蛋白快速积累的时间比AJNT-2x、H1早，且快速积累的时间长，导致AJNT-4x的蛋白含量最高。9个谷蛋白亚家族基因在花后5 d都有表达，随着胚乳的发育，各谷蛋白基因表达量逐渐升高、再逐渐下降并趋于稳定，不同的亚家族谷蛋白基因在相同的时间表达丰度不同；除GluB-1和GluB-4在H1中的表达峰值高于AJNT-4x和AJNT-2x，其他7个谷蛋白基因在H1中的表达峰值均远远低于AJNT-4x和AJNT-2x，差异显著；AJNT-4x的9个谷蛋白基因在花后30 d表达量均最高。上述结果为多倍体水稻优质育种提供理论依据。

关键词: 水稻, 四倍体, 胚乳蛋白, 谷蛋白基因

Abstract: In order to explore the relationship between different genetic composition and quality of the high-yield tetraploid rice,and the reasons for the changes in protein content,the national high-quality varieties Meixiangzhan 2(MXZ2) and Xiangyaxiangzhan(XYXZ),Aijiaonante diploid(AJNT-2x),high-yield autotetraploid Aijiaonante(AJNT-4x),and neo-tetraploid Huaduo 1(H1) were used as the test materials. The endosperm protein content was determined by BCA method,the quality analysis was performed by the near-infrared grain analyzer,the amino acid content was determined by the automatic amino acid analyzer,and the expression difference of the glutelin genes was analyzed by qRT-PCR. The results were as follows:The protein contents of brown rice and polished rice of AJNT-4x were respectively for 182.70 and 115.70 mg/g,which was much higher than that of MXZ2,XYXZ,and AJNT-2x,and the nutritional quality of AJNT-4x was greatly enhanced;the gel consistency of AJNT-4x was only for 44.70 mm,which was much lower than that of MXZ2,XYXZ,and AJNT-2x,and the difference was significant,and eating quality of AJNT-4x was significantly reduced. The protein content of polished rice of H1 was the lowest of the samples,which was only for 71.30 mg/g,but its gel consistency was for 112.70 mm,which was between the national high-quality varieties MXZ2 and XYXZ.At 30 days after flowering, the total contents of 17 amino acids and essential amino acids of AJNT-4x were higher than those of AJNT-2x and H1,and which of H1 was the lowest;the total content of 17 amino acids for H1 reached a peak at 15 days after flowering,while the total content of 17 amino acids for AJNT-4x and AJNT-2x reached a peak at 20 days after flowering,the accumulation of amino acid content stopped prematurely in H1, which resulted the lowest total content of 17 amino acids in H1 at mature period.During the whole endosperm development process,the accumulation trend of the four storage proteins was basically the same in AJNT-4x,H1,and AJNT-2x,but there were differences in the rapid accumulation time;rapid accumulation of glutelin in AJNT-4x was earlier than that in AJNT-2x and H1,and the rapid accumulation time in AJNT-4x was longer than those in AJNT-2x and H1,which resulted in highest protein content in AJNT-4x. The nine glutelin subfamily genes were expressed at 5 days after flowering. With the development of the endosperm,the expression of each glutelin gene gradually increased,then gradually decreased and tended to stabilization. Different glutelin genes had different expression abundances at the same time;except the expression peaks of GluB-1 and GluB-4 were higher in H1 than those in AJNT-4x and AJNT-2x,the expression peaks of the other seven glutelin genes were much lower in H1 than those in AJNT-4x and AJNT-2x,and the difference was significant;the expression quantities of nine glutelin genes in AJNT-4x were all the highest in the three materials at 30 days after flowering. The above results provided a theoretical basis for high-quality breeding of rice polyploidy.

Key words: Rice, Tetraploid, Endosperm protein, Glutelin gene

中图分类号:

Q78
S511.03

隆艳喜, 罗雁茹, 竺正航, 廖芷依, 王兰. 四倍体水稻蛋白质含量和谷蛋白基因表达研究[J]. 华北农学报, 2021, 36(6): 78-87. doi: 10.7668/hbnxb.20192375.

LONG Yanxi, LUO Yanru, ZHU Zhenghang, LIAO Zhiyi, WANG Lan. Research on Protein Content and Glutelin Gene Expression in Tetroploid Rice[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(6): 78-87. doi: 10.7668/hbnxb.20192375.

http://www.hbnxb.net/CN/Y2021/V36/I6/78

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