紧穗野生稻组培苗的快繁与利用研究

Study on Rapid Propagation and Utilization of Tissue Culture Seedlings of Oryza eichingeri

获得优质愈伤组织和高分化率的组培苗是离体保存和遗传转化的基础。本试验以紧穗野生稻的根为外植体,以MS和N6为基本培养基,通过添加不同浓度激素,组配不同的愈伤诱导和分化培养基,以建立适合紧穗野生稻的快繁体系。结果表明,以MS为基本培养基,2,4-D添加量少,稻根的出愈率不高,但添加量过高又抑制出愈率,以MS+11mg/L2,4-D的出愈率最高,MS+5mg/L2,4-D的愈伤增殖较好;以N6为基本培养基的愈伤诱导效果不如MS培养基。愈伤在MS+5mg/L6-BA+0.4mg/LNAA培养基中分化率最高,丛生芽多;6-BA+IAA组合的分化效果远不如6-BA+NAA好。以MS+2mg/L6-BA+0.2mg/LNAA+3mg/LTDZ作为壮苗培养基效果较理想,苗粗壮、油绿。生根培养以MS+NAA组合根露白早、主根多,MS+IAA组合远不如MS+NAA组合生根效果好;最佳生根培养基为MS+0.5mg/LNAA。利用紧穗野生稻组培苗与栽培稻配组进行远缘杂交,获得F1代杂交种子,并调查了其农艺性状表现,表明紧穗野生稻与栽培稻间具有较强的杂交优势。

Obtaining high-quality callus and high-differentiation-rate tissue culture seedlings are the base of in vitro preservation and genetic transformation. In the experiment, roots of Oryza eichingeri were used as explants. By adding different concentrations of hormones in MS and N6 medium, different callus induction and differentiation media were prepared to establish the fast propagation system suitable for Oryza eichingeri. The results showed that adding a small amount of 2,4-D in MS medium led to low callus rate of rice root, but high addition amount inhibited callus rate. When the addition amount of 2,4-D was 11 mg/L , the callus rate was the highest, and the callus proliferation was better when adding 5 mg/L 2,4-D. The callus induction effect of N6 medium was not as good as that of MS. The differentiation rate of callus in MS medium with 5 mg/L 6-BA and 0.4 mg/L NAA was the highest, which had more multiple shoots. The differentiation effect of 6-BA and IAA combination was far less than 6-BA and NAA combination. MS medium with 2 mg/L 6-BA, 0.2 mg/L NAA and 3 mg/L TDZ was the better seedling culture medium with ideal effect of thick stems and green leaves. MS and NAA combination as rooting culture medium showed white early and more taproot, so the rooting effect was far better than that of MS and IAA combination. Therefore, the best rooting medium was MS with 0.5 mg/L NAA. Distant hybridization experiment was conducted by the combination of tissue culture seedlings of Oryza eichingeri and cultivated rice in field. Then the F 1 hybrid seeds were obtained and their agronomic characters were investigated, showing strong hybridization advantage.