Tan Meimei, Chen Ruoxin, Chen Xingran, Shahid Muhammad Qasim, Liu Xiangdong, Wu Jinwen
State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, South China Agricultural University, Guangzhou 510642, China.
Base Bank of Lingnan Rice Germplasm Resources, South China Agricultural University, Guangzhou 510642, China.
Plants (Basel). 2023 Aug 20;12(16):3001. doi: 10.3390/plants12163001.
Wall is a potential genetic resource for rice breeding; however, its distant genome limits its crossing ability with cultivated rice. The interspecific hybridization of and cultivated rice, establishment of its tissue culture, and induction of polyploidy are ways to improve 's poor crossability. We developed an interspecific hybrid and studied its reproductive pollen development process in this work, and the results showed that abortive pollens (81.94%) and embryo sac abnormalities (91.04%) were the key causes of its high sterility. In order to induce callus formation in interspecific hybrid explants, two different culture media, namely Chu's N-6 medium (N6) and 1/2 Murashig and Skoog medium (1/2 MS), were employed. Additionally, two plant growth regulators (PGRs), namely 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BA), along with L-proline (Pro) and acid hydrolyzed casein, were utilized in the experiment. The optimal N6 medium, supplemented with 2.0 mg·L 2,4-D, produced the highest induction rate (80.56 ± 5.44)%. For callus differentiation and proliferation, the MS medium supplemented with 2.0 mg·L BA + 0.2 mg·L NAA produced the highest differentiation rate (75.00 ± 4.97)% and seedling emergence ratio (28.97 ± 4.67)%. The optimal combination for seedling rooting was the 1/2 MS medium supplemented with 2.0 mg L NAA and 0.2 mg L BA, which produced an average of 13.95 roots per plant. For polyploidy induction in the interspecific hybrid, the concentration of colchicine treatment at 400 mg·L for three days is an ideal protocol. We devised tissue culture and interspecific hybrid polyploidy induction to overcome ' poor crossability and introduce its favorable features into cultivated rice.
Wall是水稻育种的潜在遗传资源;然而,其亲缘关系较远的基因组限制了它与栽培稻的杂交能力。Wall与栽培稻的种间杂交、组织培养的建立以及多倍体的诱导是改善Wall较差杂交亲和性的方法。在本研究中,我们培育了一个种间杂种并研究了其生殖花粉发育过程,结果表明败育花粉(81.94%)和胚囊异常(91.04%)是其高度不育的关键原因。为了诱导种间杂种外植体形成愈伤组织,使用了两种不同的培养基,即朱的N-6培养基(N6)和1/2 Murashig和Skoog培养基(1/2 MS)。此外,实验中还使用了两种植物生长调节剂(PGR),即2,4-二氯苯氧乙酸(2,4-D)和6-苄基腺嘌呤(BA),以及L-脯氨酸(Pro)和酸水解酪蛋白。添加2.0 mg·L 2,4-D的最佳N6培养基产生了最高的诱导率(80.56 ± 5.44)%。对于愈伤组织的分化和增殖,添加2.0 mg·L BA + 0.2 mg·L NAA的MS培养基产生了最高的分化率(75.00 ± 4.97)%和出苗率(28.97 ± 4.67)%。幼苗生根的最佳组合是添加2.0 mg L NAA和0.2 mg L BA的1/2 MS培养基,平均每株产生13.95条根。对于种间杂种的多倍体诱导,400 mg·L秋水仙碱处理三天的浓度是理想的方案。我们设计了组织培养和种间杂种多倍体诱导,以克服Wall较差的杂交亲和性,并将其优良特性引入栽培稻。