Li Huanhuan, Wang Hang, Guan Lianke, Li Zihui, Wang Hua, Luo Jie
College of Horticulture and Forestry Science, Hubei Engineering Technology Research Center for Forestry Information, Huazhong Agricultural University, Wuhan 430070, China.
Life (Basel). 2023 Sep 11;13(9):1896. doi: 10.3390/life13091896.
A series of tissue culture regeneration protocols were conducted on gray poplar () to select the most efficient callus induction medium, adventitious shoot differentiation medium, shoot elongation medium and rooting medium, which laid the foundation for the optimization of genetic transformation technology for gray poplar. The results showed that the Woody Plant Medium (WPM) supplemented with 0.10 mg L kinetin (KT) and 1.00 mg L 2,4-dichlorophenoxyacetic acid (2,4-D) was the most suitable medium for callus induction. The callus induction rates of different tissues were greater than 85.7%. The optimal adventitious shoot differentiation medium was the WPM supplemented with 0.02 mg L thidiazuron (TDZ), and the adventitious shoot differentiation rates of young tissues were 22.2-41.9%. The optimal direct differentiation medium was the Murashige and Skoog (MS) medium supplemented with 0.20 mg L 6-benzylaminopurine (6-BA), 0.10 mg L indole butyric acid (IBA) and 0.001 mg L TDZ, and the differentiation rate of adventitious shoots was greater than 94%. The best shoot elongation medium for adventitious shoots was the MS medium with 0.10 mg L naphthylacetic acid (NAA). After 45 days of cultivation in the MS medium with 0.10 mg L NAA, the average plant height was 1.8 cm, and the average number of elongated adventitious shoots was 11 per explant. The 1/2 MS medium with 0.10 mg L NAA showed the best performance for rooting, and later, shoot growth. The direct shoot induction pathway can induce adventitious shoots much faster than the indirect adventitious shoot induction pathway can, and the time cost via the direct adventitious shoot induction pathway can be shortened by 2-6 weeks compared to that of the indirect shoot induction pathway.
对灰杨（）进行了一系列组织培养再生方案，以筛选出最有效的愈伤组织诱导培养基、不定芽分化培养基、芽伸长培养基和生根培养基，为优化灰杨遗传转化技术奠定了基础。结果表明，添加0.10 mg L激动素（KT）和1.00 mg L 2,4-二氯苯氧乙酸（2,4-D）的木本植物培养基（WPM）是最适合愈伤组织诱导的培养基。不同组织的愈伤组织诱导率均大于85.7%。最佳不定芽分化培养基是添加0.02 mg L噻苯隆（TDZ）的WPM，幼嫩组织的不定芽分化率为22.2-41.9%。最佳直接分化培养基是添加0.20 mg L 6-苄基腺嘌呤（6-BA）、0.10 mg L吲哚丁酸（IBA）和0.001 mg L TDZ的Murashige和Skoog（MS）培养基，不定芽分化率大于94%。不定芽最佳芽伸长培养基是添加0.10 mg L萘乙酸（NAA）的MS培养基。在添加0.10 mg L NAA的MS培养基中培养45天后，平均株高为1.8 cm，每个外植体平均伸长不定芽数为11个。添加0.10 mg L NAA的1/2 MS培养基在生根及后续芽生长方面表现最佳。直接芽诱导途径比间接不定芽诱导途径能更快地诱导不定芽，与间接芽诱导途径相比，通过直接不定芽诱导途径的时间成本可缩短2-6周。
