Relationship between embryonic callus induction from leaves and endogenous hormone content in loquat (Eriobotrya japonica Lindl.)
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摘要: 为揭示胚性愈伤组织发生过程中内源激素的变化规律，本研究以枇杷叶片为实验材料，通过诱导胚性愈伤组织获得体细胞胚，采用高效液相色谱法测定枇杷叶片及愈伤组织的赤霉素（GA3）、生长素（IAA）、脱落酸（ABA）和玉米素（ZT）4种内源激素的含量，探讨胚性愈伤组织发育过程中4种内源激素的动态变化。结果显示，不同成熟度叶片内IAA/ZT比值对胚性愈伤组织的发生有正效应，而GA3/IAA的比值具有负效应。胚性愈伤组织的发生需要较低含量的GA3及高含量的IAA和ABA，IAA/ZT比值高有利于胚性愈伤组织形成，培养后期及时添加一定量的外源激素有利于胚性保持。本研究可为离体培养时选择外植体、添加外源激素及控制继代时间提供理论指导，并为快速获得枇杷胚性材料、开展基因转化研究奠定基础。Abstract: To understand the relationship between endogenous hormone content during embryonic callus induction, somatic embryos were obtained by embryonic callus induction from loquat leaves. The content of four endogenous hormones, gibberellic acid (GA3), auxin (IAA), abscisic acid (ABA) and zeatin (ZT), were analyzed by high performance liquid chromatography (HPLC). The dynamic changes of these endogenous hormones in embryonic calli were then explored. Results suggested that the ratio of IAA/ZT in leaves had a positive effect on embryonic callus induction, whereas GA3/IAA had a negative effect. Low content of GA3 and high content of IAA and ABA were required for embryonic calli induction, and a high ratio of IAA/ZT was advantageous to their formation. The timely addition of exogenous hormones in the late period of subculture was helpful for maintaining the embryo. This research provides a theoretical direction for explant selection, extraneous hormone addition to calli subculture, and successive transfer time selection, as well as a foundation for efficiently obtaining embryonic materials and developing the study of gene transformation.
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Keywords:
- Loquat /
- Leaf /
- Embryonic callus /
- Endogenous hormone /
- Somatic embryo
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[1] Chen FX, Liu XH, Chen LS. Developmental changes in pulp organic acid concentration and activities of acid-metabolizing enzymes during the fruit development of two loquat (Eriobotrya japonica Lindl.) cultivars differing in fruit acidity[J]. Food Chem, 2009, 114:657-664.
[2] 陈小飞, 萧浪涛, 鲁旭东, 童建华. 石刁柏胚性细胞诱导过程中的内源激素和多胺含量变化[J]. 植物生理学通讯, 2006, 42(5):826-830. Chen XF, Xiao LT, Lu XD, Tong JH. Changes in endogenous phytohormone and polyamine contents during embryogenic cells induction of Asparagus officinalis L.[J]. Plant Physiology Communications, 2006, 42(5):826-830.
[3] 吴延军, 谢鸣, 蒋桂华, 陈俊伟, 吴江, 张上隆. 枇杷成熟子叶及叶片不定芽再生研究[J]. 林业科学, 2007, 43(1):107-110. Wu YJ, Xie M, Jiang GH, Chen JW, Wu J, Zhang SL. Adventitious shoot regeneration from mature cotyledons and leaves of loquat (Eriobotrya japonica)[J]. Scientia Silvae Sinicae, 2007, 43(1):107-110.
[4] 王芳, 吕洪飞, 徐红霞, 杨勇, 严其成, 汪志平. 枇杷幼叶组织培养与植株再生研究[J]. 生物技术通讯, 2011, 22(1):53-56. Wang F, Lü HF, Xu HX, Yang Y, Yan QC, Wang ZP. Plantlet regeneration from leaves of Eriobotrya japonica Lindl[J]. Letters in Biotechnology, 2011, 22(1):53-56.
[5] 贾小霞, 王汉宁, 孔维萍, 梁慧光, 张金文. 玉米幼胚和成熟胚愈伤组织诱导过程中内源激素的比较[J]. 核农学报, 2009, 24(4):555-560. Jia XX, Wang HN, Kong WP, Liang HG, Zhang JW. Comparison of dynamic changes of endogenous hormones in callus between mature and immature embryos of maize[J]. Journal of Nuclear Agricultural Sciences, 2009, 24(4):555-560.
[6] 吴顺, 萧浪涛, 刘清, 沈革志, 王若仲. 水稻愈伤组织中内源激素水平与遗传转化的关系[J]. 核农学报, 2009, 23(2):257-261. Wu S, Xiao LT, Liu Q, Shen GZ, Wang RZ. Relationship between endogenous hormones in rice calli and genetic transformation[J]. Journal of Nuclear Agricultural Scien-ces, 2009, 23(2):257-261.
[7] 田春英, 邵建柱, 刘莹, 徐继忠. 红富士苹果叶片不定芽再生中激素、多胺和NO含量的变化[J]. 园艺学报, 2010, 37(9):1403-1408. Tian CY, Shao JZ, Liu Y, Xu JZ. Change of hormones, polyamines and NO content during regeneration of adventitious buds from in vitro leaves of Red Fuji apple[J]. Acta Horticulturae Sinica, 2010, 37(9):1403-1408.
[8] 丁广文, 李华, 何桥. 植物内源激素检测技术的研究进展[J]. 基因组学与应用生物学, 2016, 35(3):1258-1265. Ding GW, Li H, He Q. Progress in study of endogenous hormones detection methods in plant[J]. Genomics and Applied Biology, 2016, 35(3):1258-1265.
[9] 胡晓辉, 江玉萍, 苗华荣, 石运庆, 陈静. 高效液相色谱法测定花生内源激素的改进研究[J]. 花生学报, 2014, 43(4):50-53. Hu XH, Jiang YP, Miao HR, Shi YQ, Chen J. Study on the improvement of HPLC method for analyzing endogenous hormone in peanut[J]. Journal of Peanut Science, 2014, 43(4):50-53.
[10] 未晓巍, 勾畅, 徐民泽, 徐洪伟, 周晓馥. 高效液相色谱法测定玉米内源激素方法的改进研究[J]. 玉米科学, 2013, 21(3):144-148. Wei XW, Gou C, Xu MZ, Xu HW, Zhou XF. Study on the improvement of HPLC method for analyzing endogenous hormone in maize[J]. Journal of Maize Sciences, 2013, 21(3):144-148.
[11] 张媛媛, 马晖玲, 俞玲, 赵媛. 草地早熟禾愈伤组织诱导和分化中内源激素水平分析[J]. 甘肃农业大学学报, 2013, 48(2):74-79. Zhang YY, Ma HL, Yu L, Zhao Y. Tissue culture and endogenous hormones change of Poa Pratensis[J]. Journal of Gansu Agricultural University, 2013, 48(2):74-79.
[12] 马海燕, 王美丽, 张振文. 葡萄新梢生长过程中内源激素含量的动态变化[J]. 西北农业学报, 2007, 16(4):177-179. Ma HY, Wang ML, Zhang ZW. Changes of endogenous hormones in the shoots of grapevine[J]. Acta Agriculturae Boreali-occidentalis Sinica, 2007, 16(4):177-179.
[13] 曾庆钱, 陈厚彬, 鲁才浩, 李建国. HPLC测定荔枝不同器官中内源激素流程的优化[J]. 果树学报, 2006, 23(1):145-148. Zeng QQ, Chen HB, Lu CH, Li JG. An optimized HPLC procedure for analyzing endogenous hormones in different organs of litchi[J]. Journal of Fruit Science, 2006, 23(1):145-148.
[14] 周红玲, 赖钟雄, 郑加协, 张娟, 李永裕, 陈石. 梯度洗脱测定枇杷试管苗内源激素方法的研究[J]. 热带作物学报, 2010, 31(12):2292-2296. Zhou HL, Lai ZX, Zheng JX, Zhang J, Li YY, Chen S. Determination of intrinsic hormones in plantlets in vitro of loquat cultivar by HPLC with gradient elution[J]. Chinese Journal of Tropical Crops, 2010, 31(12):2292-2296.
[15] 高庆玉, 闫超, 张丙秀, 王振武, 王雪娇, 张宇. 树莓叶片生长过程中内源激素含量变化初步研究[J]. 湖北农业科学, 2015, 54(24):6280-6283. Gao QY, Yan C, Zhang BX, Wang ZW, Wang XJ, Zhang Y. Changes of endogenous hormone content during the growth of raspberry leaves[J]. Hubei Agricultural Scien-ces, 2015, 54(24):6280-6283.
[16] Rai MK, Shekhawat NS, Harish, Gupta AK, Phulwaria M, Ram K, Jaiswal U. The role of abscisic acid in plant tissue culture:a review of recent progress[J]. Plant Cell Tissue Organ Cult, 2011, 106:179-190.
[17] Huang WL, Lee CH, Chen YR. Levels of endogenous abscisic acid and indole-3-acetic acid influence shoot organogenesis in callus cultures of rice subjected to osmotic stress[J]. Plant Cell Tissue Organ Cult, 2012, 108:257-263.
[18] 付凤玲, 冯质雷, 渠柏艳, 李晚忱. 玉米未成熟胚胚性愈伤组织诱导率与内源激素含量的关系[J]. 核农学报, 2006, 20(1):10-14. Fu FL, Feng ZL, Qu BY, Li WC. Relationships between induction rate of embryonic callus and endogenous hormones content in maize[J]. Journal of Nuclear Agricultu-ral Sciences, 2006, 20(1):10-14.
[19] 郭敏敏, 王清连, 胡根海. 利用高效液相色谱法分离和测定棉花组织培养过程中4种内源激素[J]. 生物技术通讯, 2009, 20(2):213-216. Guo MM, Wang QL, Hu GH. Separation and determination of four plant hormones during somatic embryogenesis of Gossypium hirsutum L. with HPLC[J]. Letters in Biotechnology, 2009, 20(2):213-216.
[20] 刘艳君, 张超, 杨静慧, 李冰, 刘婷, 秦艳筠. 塔杨松散型均质胚性愈伤组织培养体系[J]. 植物研究, 2016, 36(1):123-128. Liu YJ, Zhao C, Yang JH, Li B, Liu T, Qin YJ. Culture system on loose, homogeneous, embryogenic callus of Populos×canadensis Moench ‘Tower’[J]. Bulletin of Botanical Research, 2016, 36(1):123-128.
[21] 洪森荣, 尹明华, 王艾平. 江西铅山红芽芋胚性愈伤组织的包埋玻璃化超低温保存[J]. 植物科学学报, 2014, 32(1):80-87. Hong SR, Yin MH, Wang AP. Cryopreservation of Jiangxi Yanshan red bud taro embryogenic calli by encapsulation-vitrification[J]. Plant Science Journal, 2014, 32(1):80-87.
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