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以4份高粱不育系和5种类型苏丹草为亲本,按照NCⅡ设计配制成20个杂交组合,分析各组合及亲本的表型值和中亲及超亲优势并筛选出8个优势强的组合为试材,利用cDNA-AFLP技术,分析杂种与亲本苗期叶片基因差异表达类型与主要产量性状的杂种表现及杂种优势的关系。研究表明:(1)12对引物共扩增出315条TDFs,杂种与亲本间基因表达类型有:单亲表达一致一型(P1F1型)和二型(P2F1型)、杂种特异表达类型(F1型)、单亲表达沉默一型(P1型)和二型(P2型)、双亲共沉默类型(P1P2型)和杂种亲本表达一致型(P1F1P2型)7种。(2)在差异展示类型与产量构成因素的相关分析中,有效分蘖数与P1F1型(0.726**)呈极显著正相关,单株鲜重与P1P2型(0.659*)、叶长与P2型(0.647*)呈显著正相关,成株期叶片数与F1型(-0.81**)呈极显著负相关。在与中亲优势相关分析中发现,单株鲜重与P1(0.695*)、P2(0.637*)呈显著正相关,单株鲜重与P1F1P2型(0.743**)呈极显著正相关,叶宽与P1P2型(-0.619*)呈显著负相关。在与超亲优势进行相关分析后发现,穗长与P2F1型(0.732**)呈极显著正相关,叶宽与P2F1型(-0.731**)以及P1P2型(-0.731**)呈极显著负相关。(3)差异展示类型P1F1、P2F1、P1和P2是显性效应类型,共占总检测的91.4%。差异展示类型F1和P1P2表现超显性,共占总检测的4.8%,说明各个性状的杂种表现主要受到的是(超)显性效应影响。(4)对8个与高丹草杂种优势相关的TDFs进行回收及BLAST分析均得到同源核苷酸,并且找到7个同源蛋白,这些蛋白质在控制植物生长发育方面具有重要作用。(5)将克隆测序获得差异片段的核苷酸序列,采用半定量RT-PCR进行了验证。本研究为进一步揭示高丹草杂种优势的分子机制和提高高丹草强优势组合的筛选效率以及种质资源的创建提供依据。
Four sorghum CMS lines and five types of sudangrass were used as parents, and 20 hybrid combinations were designed according to NCⅡ. The phenotypic values, mid-progeny and super-parents of each combination and their parents were analyzed and eight strong And the relationship between heterosis and heterosis was analyzed by cDNA-AFLP technique. The results showed that: (1) A total of 315 TDFs were amplified by 12 pairs of primers. The gene expression patterns between F1 hybrids and parents were P1F1 type and P2F1 type, F1 type ), Single parent expressed silence type 1 (type P1) and type 2 (type P2), parents co-silence type (P1P2 type) and hybrid parent expression consistent type (P1F1P2 type) 7 species. (2) Correlation analysis showed that there was a significant positive correlation between the effective tiller number and P1F1 type (0.726 **). The correlation between the fresh weight and P1P2 type (0.659 *), leaf length and P2 type (0.647 *). There was a significant negative correlation between the number of leaves at adult stage and F1 (-0.81 **). Correlation analysis showed that the fresh weight per plant was positively correlated with P1 (0.695 *) and P2 (0.637 *). The fresh weight per plant was significantly and positively correlated with P1F1P2 (0.743 **) The width was negatively correlated with P1P2 (-0.619 *). Correlation analysis showed that there was a significant positive correlation between spike length and P2F1 (0.732 **). Leaf width was significantly different from that of P2F1 (-0.731 **) and P1P2 (-0.731 **) Negative correlation. (3) Difference display types P1F1, P2F1, P1 and P2 are dominant effect types, accounting for 91.4% of the total testing. Difference display types F1 and P1P2 showed overdominance, accounting for 4.8% of the total, indicating that the hybrid performance of each trait was mainly affected by (super) dominance effect. (4) Eight homologous nucleotides were obtained from TDFs related to Gordian hybrid vigor and BLAST analysis, and seven homologous proteins were identified. These proteins play an important role in controlling plant growth and development. (5) The nucleotide sequence of the difference fragment was cloned and sequenced, and verified by semi-quantitative RT-PCR. This study provides the basis for further revealing the molecular mechanism of Sorghum sudanense heterosis and improving the screening efficiency of Sorghum sudanense strong-dominant combination and the establishment of germplasm resources.