摘要
opaque-2(o2)基因可以显著提高玉米胚乳中的赖氨酸和色氨酸含量,胚乳修饰基因可改善胚乳物理性状,增加籽粒硬质度、粒重、透明度。本研究利用与opaque-2(o2)基因紧密连锁的分子标记phi057,检测QPM 群体中群13、中群14 内o2 基因的频率分布,发现中群13、中群14 在o2 基因位点的等位基因频率发生了很大变化,说明定期监测群体o2o2 基因型频率的必要性。利用phi057 分子标记检测CB、CD、CA、R 高代自交系,发现某些高代系在o2 基因位点上没有纯合,通过分子标记进行o2 基因的检测,可以提高选择的准确性。
将现有的优良普通玉米自交系转育成QPM,是丰富优质蛋白玉米遗传基础的有效途径。本研究通过优化DNA 提取技术、SSR、AFLP 技术流程,建立了一整套进行QPM的分子标记辅助选择体系。在将50 个普通玉米自交系转育成QPM 自交系的过程中,利用分子标记辅助选择O2o2 基因型,结果表明是十分有效的。利用与o2 基因紧密连锁的分子标记对CA335 和黄早四回交群体、CA335 和178 回交群体的BC1F1 和BC2F1的前景选择,表明利用分子标记能够有效地进行QPM o2 基因的前景选择;利用AFLP 分子标记进行QPM 回交群体BC1F1 和BC2F1的背景选择,表明利用AFLP 分子标记辅助选择使回交二代比回交一代与轮回亲本的相似性显著增加, 使未纯合位点进一步纯合,从而加快QPM 育种进程,为分子标记辅助选择QPM 提供重要理论依据。
opaque-2(o2)gene increases the lysine and tryptophan contents sinificantly in maizeendosperm. Modifying genes could alter the kernel phenotype including kernel horniness,kernel weightiness and kernel transparency. In this study, the SSR primer phi057 that linkedtightly with opaque-2 (o2) was used to detect o2o2 genotype in the QPM(Quality ProteinMaize) populations CPop13, and CPop14, and it was revealed that the frequency of o2 genehad shifted significantly in the two populations. The MAS was used to detect the o2o2genotype in CA, CD, CB, and R series lines. It was indicated that homozygous o2o2 genotypein QPM inbred lines could not be ensured with conventional breeding efforts.
Germplasm background is constraint in QPM breeding efforts. In the current study anefficient technique of marker assisted selection for QPM was established based on theoptimized DNA extraction, technique program of SSR (Simple Sequence Repeat) andAFLP(Amplified Fragment Length Polymorphism). In order to enhance the QPM germplasmbasis and convert 50 elite normal lines into QPM, the SSR marker Phi057 was used to detecto2 gene in backcross generations, and it proved to be a powerful tool in the efforts. The SSRprimer phi057 was used to detect o2 gene in BC1F1 and BC2F1 of the crosses(CA335×X178) , (CA335×HZ4). It was proved that the marker used to foreground selectionis effective. The AFLP primers were used to detect background in BC1F1 and BC2F1 of thecrosses. It was proved that the similarity of BC2F1 increases more apparently than similarityof BC1F1, and more heterozygous loci tend to homozygous. Molecular Markers AssistedSelection (MAS) can expedites breeding program. It offers important theoretical method forMolecular Markers Assisted Selection of QPM.
引文
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