野生二粒小麦NAM-B1基因导入川农16后的表达情况及对品质的影响
发布时间：2018-11-05 18:31
【摘要】：野生二粒小麦是普通小麦A、B染色体组的供体祖先种,具有籽粒蛋白质含量高且籽粒微量元素含量高等特点。现代栽培小麦具有较高的产量,但品质亟待提高。NAM-B1基因在野生二粒小麦中普遍存在,在生育后期表达,一方面加速植株衰老,另一方面促进营养器官中的N、Fe、Zn等营养物质向发育中的籽粒转运。然而在现代栽培小麦中,NAM-B1基因或是被删除,或是由于在编码区中第11个碱基位点发生T碱基插入,导致移码突变而丧失功能。本实验通过对野生二粒小麦D1和D97与普通小麦川农16杂交F10代进行NAM-B1基因克隆和表达量测定,籽粒收获后测定籽粒蛋白质含量和铁、锌含量,探究NAM-B1基因导入川农16后的序列变异和表达情况及该基因对营养品质的影响。主要研究结果如下：D1和D97携带表达型NAM-B1基因,其序列相比功能型NAM-B1基因序列DQ869673只有一个碱基的同义突变；川农16没有对应PCR扩增产物,说明该基因被删除。27份杂交Flo代株系中12份材料克隆结果与川农16一致,系缺失型；12份材料克隆结果与D1和D97一致,系野生型;3份材料NAM-B1基因序列存在编码区中第11个碱基位点发生T碱基插入而导致的移码突变,系无功能的突变型。开花期NAM-B1基因表达量处于较低水平,范围是actin基因表达量的1/22—1/6,且不同材料间差异不显著；开花15天后表达量较高,范围是actin基因表达量的1/4.5—1/1.5,不同材料间表达量存在极显著差异。携带野生型NAM-B1基因的F10代材料平均籽粒蛋白质含量显著高于不携带野生型NAM-B1基因的F10代材料。12份携带野生型NAM-B1基因的F1o代材料中有4份显著高于亲本川农16,而15份不携带野生型NAM-B1基因的F10代材料中只有1份显著高于川农16。携带野生型NAM-B1基因的F1o代材料平均籽粒Fe含量显著高于不携带野生型NAM-B1基因的F10代材料,同样显著高于亲本川农16。而携带野生型NAM-B1基因的Flo代材料、不携带野生型NAM-B1基因的F10代材料以及川农16平均籽粒Zn含量差异不显著。携带和不携带野生型NAM-B1基因的F10代材料平均千粒重差异不显著,但是均显著高于亲本川农16,这很可能“归功于”我们自F2代起不断沿着高产育种方向进行选择。因为高粒重的材料存在“稀释”效应,所以单位质量籽粒中的营养物质含量只是相对含量,不能正确反映营养物质向籽粒转运的能力。计算千粒重和籽粒营养物质含量的乘积得到的每千粒营养物质含量才是绝对含量。野生型NAM-B1基因在六倍体背景下提高籽粒蛋白质绝对含量和籽粒铁绝对含量,但是对籽粒锌绝对含量的作用效果没有达到显著性水平。相关分析结果显示携带野生型NAM-B1基因的Fl0代材料各营养品质指标间相关性明显强于不携带野生型NAM-B1基因的F10代材料。籽粒营养物质相对含量和千粒重全部表现为负相关,有些达到了显著水平,相比之下,籽粒营养物质绝对含量和千粒重相关性较弱,对于携带野生型NAM-B1基因的F1o代材料还表现为正相关。因为籽粒营养物质绝对含量才真正反映向籽粒转运营养物质的能力,所以我们认为在后代中选育粒大、千粒重高的株系并不会造成品质低下的问题,通过对NAM-B1基因的利用、配合田间选种,就很有希望培育出既高产又优质的小麦新品种。此外,NAM-B1基因的表达量和各营养品质指标的相关分析还表明影响蛋白质和铁、锌含量的因素很复杂,除了NAM-B1基因之外很可能还有其它基因在调控这些生理生化过程。
[Abstract]:Wild two-grain wheat is the donor ancestor of wheat A and B chromosome group, and has the characteristics of high protein content and high content of trace elements in grains. The modern cultivated wheat has higher yield, but the quality needs to be improved. The NAM-B1 gene is ubiquitous in the wild two-grain wheat, which is expressed in late-growing stage. On the one hand, the plant senescence is accelerated, and on the other hand, nutrient substances such as N, Fe, Zn and the like in the vegetative organs are transferred to the grain in the development. However, in modern cultivation of wheat, the NAM-B1 gene is either deleted, or due to the insertion of a T-base at the 11th nucleotide site in the coding region, the shift mutation is lost and the function is lost. In this experiment, the cloning and expression of NAM-B1 gene were determined by crossing F10 generations of wild two wheat D1 and D97 and common wheat Chuangnong 16, and the content of protein and iron and zinc content were determined after grain harvest. To investigate the sequence variation and expression of NAM-B1 gene and the effect of the gene on nutritional quality. The main results are as follows: D1 and D97 carry the expression type NAM-B1 gene, the sequence of which is similar to that of the functional NAM-B1 gene sequence DQ869673, only one base has synonymous mutation; the Sichuan farmer 16 does not correspond to the PCR amplification product, The gene was deleted. 12 of the 27 hybrid Flo strains were consistent with the Chuangnong 16, and the results of 12 materials were consistent with D1 and D97, which were wild-type. 3 portions of the NAM-B1 gene sequence present a shift mutation resulting from the insertion of a T base at the 11th nucleotide site in the coding region, and is a non-functional mutant. The expression level of NAM-B1 gene in flowering stage was at a low level, the range was 1/ 22/ 1/ 6 of the expression of survivin gene, and the difference between different materials was not significant. The average grain protein content of F10 generation material carrying wild type NAM-B1 gene is significantly higher than that of F10 generation material carrying wild type NAM-B1 gene. Only 1 of the 10-generation materials which did not carry the wild-type NAM-B1 gene were significantly higher than that of the non-wild-type NAM-B1 gene. The average grain Fe content of F1o material carrying the wild-type NAM-B1 gene is significantly higher than that of the F10 generation material which does not carry the wild-type NAM-B1 gene, and is also significantly higher than that of the parent Sichuan farmer 16. However, the Fo material carrying the wild-type NAM-B1 gene does not carry the F10 generation material of the wild-type NAM-B1 gene and the average grain Zn content of the Sichuan farmer 16 is not significant. The average 1000-grain weight difference of the F10 generation materials carrying and without the wild-type NAM-B1 gene was not significant, but they were significantly higher than that of the parent Chuangnong 16, which was very likely to 鈥渙we to鈥，
本文编号：2312984
[Abstract]:Wild two-grain wheat is the donor ancestor of wheat A and B chromosome group, and has the characteristics of high protein content and high content of trace elements in grains. The modern cultivated wheat has higher yield, but the quality needs to be improved. The NAM-B1 gene is ubiquitous in the wild two-grain wheat, which is expressed in late-growing stage. On the one hand, the plant senescence is accelerated, and on the other hand, nutrient substances such as N, Fe, Zn and the like in the vegetative organs are transferred to the grain in the development. However, in modern cultivation of wheat, the NAM-B1 gene is either deleted, or due to the insertion of a T-base at the 11th nucleotide site in the coding region, the shift mutation is lost and the function is lost. In this experiment, the cloning and expression of NAM-B1 gene were determined by crossing F10 generations of wild two wheat D1 and D97 and common wheat Chuangnong 16, and the content of protein and iron and zinc content were determined after grain harvest. To investigate the sequence variation and expression of NAM-B1 gene and the effect of the gene on nutritional quality. The main results are as follows: D1 and D97 carry the expression type NAM-B1 gene, the sequence of which is similar to that of the functional NAM-B1 gene sequence DQ869673, only one base has synonymous mutation; the Sichuan farmer 16 does not correspond to the PCR amplification product, The gene was deleted. 12 of the 27 hybrid Flo strains were consistent with the Chuangnong 16, and the results of 12 materials were consistent with D1 and D97, which were wild-type. 3 portions of the NAM-B1 gene sequence present a shift mutation resulting from the insertion of a T base at the 11th nucleotide site in the coding region, and is a non-functional mutant. The expression level of NAM-B1 gene in flowering stage was at a low level, the range was 1/ 22/ 1/ 6 of the expression of survivin gene, and the difference between different materials was not significant. The average grain protein content of F10 generation material carrying wild type NAM-B1 gene is significantly higher than that of F10 generation material carrying wild type NAM-B1 gene. Only 1 of the 10-generation materials which did not carry the wild-type NAM-B1 gene were significantly higher than that of the non-wild-type NAM-B1 gene. The average grain Fe content of F1o material carrying the wild-type NAM-B1 gene is significantly higher than that of the F10 generation material which does not carry the wild-type NAM-B1 gene, and is also significantly higher than that of the parent Sichuan farmer 16. However, the Fo material carrying the wild-type NAM-B1 gene does not carry the F10 generation material of the wild-type NAM-B1 gene and the average grain Zn content of the Sichuan farmer 16 is not significant. The average 1000-grain weight difference of the F10 generation materials carrying and without the wild-type NAM-B1 gene was not significant, but they were significantly higher than that of the parent Chuangnong 16, which was very likely to 鈥渙we to鈥，
本文编号：2312984
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