In order to obtain haploid materials of Pinus massoniana and establish endosperm tissue culture system, different disinfectants and disinfection times of P. massoniana were screened, and the effects of different hormone ratios and different heat shock temperatures and times on callus induction were studied. The results showed that the best way to sterilize explants was 75% alcohol treatment for 30 s+2% NaClO for 15 min, the explant contamination rate was 0, the mortality rate was only 15.56%; Auxin could promote callus induction and was necessary; The seeds soaked with cytokinin could replace the cytokinin in the medium, and could obtain better effect of endosperm callus induction, 1.0 mg·mL-1 6-BA for 30 min was the best, in the medium WPM+NAA 2.0 mg·L-1+2,4-D 1.0 mg·L-1+sucrose 30 g·L-1+agar 7 g·L-1 the induction rate was as high as 87.78%; The medium DCR+NAA 2.0 mg·L-1+6-BA 1.0 mg·L-1+sucrose 30 g·L-1+agar 7 g·L-1 had the highest endosperm callus induction rate (68.75%); Heat shock treatment could improve the induction rate and quality of endosperm callus. The best heat shock at 45 ℃ under 85% relative humidity for 10 minutes, the total callus induction rate was 66.25%, and the mass callus induction reached 8.75%; The best proliferation culture was WPM+NAA 0.1 mg·L-1+6-BA 2.0 mg·L-1+sucrose 30 g·L-1+agar 7 g·L-1, and the proliferation rate was 83.33%.
YUAN Yu-hong, SUN Yan, ZHENG Yu-shan
. A Preliminary Study on Occurrence of Endosperm Callus in Pinus massoniana[J]. Subtropical Plant Science, 2021
, 50(03)
: 182
-188
.
DOI: 10.3969/j.issn.1009-7791.2021.03.004
[1] 周政贤. 中国马尾松[M]. 北京: 中国林业出版社, 2001: 14–15.
[2] 秦国峰. 马尾松培育及利用[M]. 北京: 金盾出版社, 1997: 135–180.
[3] 杨模华,李志辉,张冬林,黄振,丁贵杰. 马尾松嫩茎愈伤组织保持、增殖与不定芽分化培养[J]. 中国农学通报, 2011,27(10): 12–17.
[4] 季孔庶,王章荣,陈天华,王明庥. 马尾松扦插繁殖年龄效应及继代扦插复壮效果[J]. 浙江林学院学报, 1999,16(4): 13–17.
[5] 杨艳. 马尾松幼胚培养及幼茎愈伤组织诱导研究[D]. 长沙: 中南林业科技大学硕士学位论文, 2007.
[6] 黄振. 马尾松幼苗茎段和春梢茎段组织培养[D]. 长沙: 中南林业科技大学硕士学位论文, 2009.
[7] 唐婷婷. 马尾松优良种源桐棉松组培技术的研究[D]. 南宁: 广西大学硕士学位论文, 2015.
[8] 杨模华,张冬林,李志辉,靳小翠,丁贵杰. 马尾松幼胚体细胞胚胎发生研究[J]. 植物生理学报, 2011,47(9): 904–912.
[9] 郑武林. 马尾松与落叶松高效离体再生体系建立[D]. 南昌: 江西农业大学硕士学位论文, 2014.
[10] 薛鹰. 马尾松组织培养快速繁殖技术的研究[D]. 南宁: 广西大学硕士学位论文, 2006.
[11] 王胤,姚瑞玲. 马尾松优良种源高效组培育苗技术体系构建[J]. 北京林业大学学报, 2020,42(6): 43–51.
[12] 王胤,姚瑞玲,李慧娟,张烨. 基于外植体生理复幼的马尾松茎段芽无菌离体培养[J]. 植物生理学报, 2019,55(9): 1375–1384.
[13] 姚瑞玲,王胤,吴幼媚. 马尾松组培生根关键因子分析[J]. 广西植物, 2016,36(11): 1288–1294,1375.
[14] 姚瑞玲,王胤. 温度对马尾松组培单芽不定根发生的影响[J] .广西植物, 2016,36(11): 1282–1287.
[15] 齐力旺,韩素英,杨云龙,李怀忠,吴晓玲. 几种裸子植物胚乳愈伤组织的诱导[J]. 应用与环境生物学报, 1997,3(1): 19–22,91.
[16] 顾淑荣,朱至清,赵敬芳,陈正华. 白皮松和油松雌配子体愈伤组织的诱导和分化[J]. 植物学报, 1995,37(3): 217–221.
[17] 杜雪玲,张振霞,余如刚,符义坤. 植物组织培养中的污染成因及其预防[J]. 草业科学, 2005,22(1): 24–27.
[18] 张东方. 植物组织培养培养技术[M]. 哈尔滨: 东北林业大学出版社, 2004: 12–15.
[19] 黄健秋,卫志明. 松属树种的组织培养和原生质体培养[J]. 植物学通报, 1994,11(1): 34–42.
[20] 吴元立,严学成. 银杏成熟胚乳培养的细胞组织学观察[J]. 果树科学, 1998,15(4): 327–331.
[21] 初立业,宏波邵. 植物激素在松树离体快速繁殖中的作用[J]. 生物技术通报, 1995(1): 6–10.
[22] 吕晓波,陈力,陈香兰. 热激对低温预处理粳稻花药培养的影响[J]. 植物生理学通讯, 1991(3): 209–210.
[23] 齐力旺,杨云龙,韩素英,冯建成,王勇强. 樟子松胚乳单倍体愈伤组织发生与分化的研究[J]. 林业科技通讯, 1996(8): 17–18,25.
[24] 王敬驹,陆文梁,匡柏健. 小黑麦杂种胚乳的离体培养研究[J]. 植物学报, 1982,24(5): 420–425,500.
