期 刊 ：植物遗传资源学报 2016年 17卷 4期
关键词：高丹草；cDNA-AFLP；半定量RT-PCR；杂种优势；
Keywords:Sorghum, cDNA-AFLP, semi-quantitative RT-PCR, heterosis,
摘 要 ：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，分析各杂交组合及亲本的表型值和中亲优势及超亲优势并筛选出8个杂种优势强的杂交组合为材料，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，高丹草杂种与亲本间基因表达类型有：单亲表达一致一型 (P1F1型)、单亲表达一致二型 (P2F1型)、杂种特异表达类型 (F1型)、单亲表达沉默一型 (P1型)、单亲表达沉默二型 (P2型)、双亲共沉默类型 (P1P2型)和杂种亲本表达一致型（P1F1P2型）七种。（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、单株鲜重与P1P2型呈显著正相关，成株期叶片数与F1型、叶长与P2型呈显著负相关。在与中亲优势相关分析中发现，穗长与P2F1型、单株鲜重与P1和P2以及P1F1P2型呈显著正相关，叶宽与P1P2型呈显著负相关。在与超亲优势进行相关分析后发现，穗长与P2F1型呈显著正相关，叶宽与P2F1与P1P2型呈显著负相关。（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）通过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。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。
Abstract:Four copies of the sorghum sterile lines and five types of Sudan grass for parents, according to the NC Ⅱ design，were formulated to 20 hybrid combinations. Eight hybrid combinations of strong heterosis were selected with evaluation of the phenotypic value and the mid-parent advantage. heterobeltiosis Using cDNA-AFLP technology, differentially expressed genes from hybrid and parental seedling leafwere analyzed and correlated with main yield traits of hybrid performance and the heterosis. Research shows that: (1) 12 pairs of primers amplified a total of 315 TDFs, the gene expression types of sorghum-sudan grass were classified into 7 catagries: single parent expression of the same type (P1F1 type) , single parent expression with type 2 (p2f1 type), hybrid specific expression type (F1), single parent expression silence type 1(P1), a single expression silence type 2 (P2), and parents silent type (P1P2 type) and hybrid- parents joint expression type (P1F1P2 type). (2) the correlation analysis of yield components showed significant positive correlation between effective tiller number and P1F1 type, fresh weight per plant and P1P2 type; it showed significant negative correlation between leaf number and F1 type, leaf length and P2 type. the correlation analysis between mid-parent heterosis and gene types showed significant positive correlation between the spike length and the P2F1 type, fresh weight per plant with P1, P2 and P1F1P2; significant negative correlation between leaf width and P1P2. After correlation analysis with the super parent heterosis, the spike length was significantly positively correlated with P2F1, and the leaf width was significantly negatively correlated with P2F1 and P1P2. (3) differential display types of P1F1, P2F1, P1 and P2 are dominant effects, which are 91.4% of the total. Differential Display type F1 and P1P2 performance super dominance, acount for 4.8% . It indicated that the hybrid performance of each trait was mainly influenced by the (super) dominant effect. (4) 8 TDFS associated heterosis were retrieved and found homologous nucleotides with BLAST analysis, and seven homologous proteins were found important in the control of plant growth and development. (5) the nucleotide sequence was obtained by cloning and sequencing, and semi quantitative RT-PCR was used to verify the nucleotide sequence. The molecular mechanisms of heterosis were revealed, and the selecting efficiency should be improved in screening strong advantage combinationsand providing basis for the creation of germplasm resources in Sorghum-sudan grass.