黄瓜10个基因型材料外植体内源激素水平及比例对其离体再生的影响

doi:10.16420/j.issn.0513-353x.2020-0855

摘要/Abstract

摘要：

以10个不同基因型黄瓜（Cucumis sativus L.）子叶节外植体为材料,研究外植体内源激素水平、诱导分化阶段不同外源激素及配比对外植体分化率、不定芽数量和形态的影响,揭示影响不同基因型黄瓜离体再生活力的关键激素因素,并进一步优化黄瓜再生体系。结果表明：MS培养基不添加植物生长调节剂时,10个材料的分化率差异显著,内源CTK水平与分化率呈显著正相关,相关系数分别为0.4544（培养0 d）和0.6110（培养14 d）。MS培养基添加外源1.5 mg · L^-16-BA后,不同基因型的分化率从5.95% ~ 20.45%提高到了42.39% ~ 80.43%。不定芽形态观察发现：外植体再生过程形成了两种不同类型的不定芽：Ⅰ型芽为正常芽,叶片舒展,茎秆明显;Ⅱ型芽为畸形芽,有叶片形态,但无明显的茎秆结构。不同基因型材料分化形成Ⅰ型和Ⅱ型不定芽的比例差异显著,Ⅰ型/Ⅱ型不定芽比值高的基因型最终再生苗率显著高于比值低的基因型。相关性分析显示不同材料中产生Ⅰ型/Ⅱ型外植体比例与其IAA/ABA比值的相关性最高（R²= 0.4214）,Ⅰ型/Ⅱ型不定芽比值与再生苗率呈显著正相关（R²= 0.5623）。对Ⅰ型/Ⅱ型外植体比值低（0.52和0.46）、不定芽比值低且再生苗率较低的基因型C5和C6,通过添加不同比例外源NAA和ABA进行诱导,发现C5和C6均在NAA/ABA比值为0.01/0.1时Ⅰ型/Ⅱ型外植体比例（1.65和1.15）、不定芽比值（1.81和1.17）和再生苗率（48.57%和53.33%）均得到显著提高。

关键词: 黄瓜, 离体再生, 激素因素, 体系优化

Abstract:

In order to reveal the key factor influencing the regeneration vigor of different cucumber （Cucumis sativus L.）and optimize the cucumber in vitro regeneration system. The effect of endogenous hormone levels and exogenous hormone treatments on the differentiation rate,the number and morphology of adventitious buds were investigated in cotyledon node explants of 10 different genotypes. The results showed that the differentiation rate of the 10 materials was significantly different in the absence of exogenous hormones,and the endogenous CTK level was positively correlated with the differentiation rate,with correlation coefficients were 0.4544 （0 d incubation）and 0.6110（14 d incubation）,respectively. The differentiation rates of different genotypes were increased from 5.95% ~ 20.45% to 42.39% ~ 80.43% by 1.5 mg · L^-16-BA treatment. Morphology observation showed there were two different types of adventitious buds,typeⅠbuds：leaf blade stretching and visible culm,typeⅡbuds：deformity bud with leaf blade and no visible culm. The proportion of the formation of typeⅠand typeⅡadventitious buds were significantly different in different genotypes. The regeneration rates of the genotypes with a high ratio of typeⅠ/Ⅱadventitious buds were significantly higher than that of the genotypes with a low ratio. Correlation analysis showed that the typeⅠ/Ⅱexplant ratio was closely correlated with the IAA/ABA ratio（R²= 0.4214）,and the ratio of typeⅠ/Ⅱadventitious buds was obviously positive correlation with the rate of regeneration seedings（R²= 0.5623）. For the genotype C5 and C6,which had a low typeⅠ/Ⅱexplant ratio（0.52 and 0.46）,adventitious buds ratio and low rate of regeneration seedings,different proportion of exogenous NAA and ABA was added to the induction medium,it was found that typeⅠ/Ⅱexplants ratio（1.65 and 1.15）,adventitious buds ratio（1.81 and 1.17）and the regeneration rate（48.57% and 53.33%）were significantly improved when the NAA/ABA ratio was 0.01/0.1.

Key words: cucumber, regeneration in vitro, hormonal factor, system optimization

中图分类号:

S642.2

王团团, 缑晨星, 夏磊, 朱拼玉, 李季, 陈劲枫. 黄瓜10个基因型材料外植体内源激素水平及比例对其离体再生的影响[J]. 园艺学报, 2021, 48(9): 1731-1742.

WANG Tuantuan, GOU Chenxing, XIA Lei, ZHU Pinyu, LI Ji, CHEN Jinfeng. Effect of Endogenous Hormone Levels and Ratios in Explants of Ten Genotypes of Cucumber on Their Regeneration in Vitro[J]. Acta Horticulturae Sinica, 2021, 48(9): 1731-1742.

图/表 11

参考文献 29

[1]	Aydin . 2013. Plant regeneration in wheat mature embryo culture. African Journal of Biotechnology, 10(70):15749-15755.
[2]	Bömer M, O'Brien J, Perez-Salamo I, Krasauskas J, Finch P, Briones A, Daudi A, Souda P, Tsui T L, Whitelegge J P, Bolwell G P, Devoto A. 2018. COI1-dependent jasmonate signaling affects growth,metabolite production and cell wall protein composition in Arabidopsis. Annals of Botany, 122(7):1117-1129.
[3]	Du Sheng-li, Wei Hui-jun, Wei Ai-min, Wang Yan-fei, Ma De-hua, Huo Zhen-rong. 2000. Effects of seedling stage,genotype and explant type on in vitro somatic organogenesis of cucumber. Tianjin Agricultural Sciences,(4):1-5. (in Chinese)
	杜胜利, 魏惠军, 魏爱民, 王艳飞, 马德华, 霍振荣. 2000. 苗龄、基因型和外植体类型对黄瓜离体器官发生的影响. 天津农业科学,(4):1-5.
[4]	Gao Z H, Zhang H Y, Cao C X, Han J, Li H, Ren Z H. 2020. QTL mapping for cucumber fruit size and shape with populations from long and round fruited inbred lines. Horticultural Plant Journal, 6(3):132-144. doi: 10.1016/j.hpj.2020.04.004 URL
[5]	Grozeva S, Velkov N. 2014. In vitro plant regeneration of two cucumber(Cucumis sativum L.) genotypes:effects of explant types and culture medium. Genetika, 46(2):485-493. doi: 10.2298/GENSR1402485G URL
[6]	Gu Rui-sheng, Jiang Xiang-ning, Guo Zhong-chen. 1999. Advances in the studies on the mechanism of plant organogenesis in vitro. Chinese Bulletin of Botany,(3):3-5. (in Chinese)
	谷瑞升, 蒋湘宁, 郭仲琛. 1999. 植物离体培养中器官发生调控机制的研究进展. 植物学通报,(3):3-5.
[7]	Hiroshi H, Takakazu M, Izumi C M, Miki Y, Kazuhiro S. 2016. Endogenous hormone levels affect the regeneration ability of callus derived from different organs in barley. Plant Physiology & Biochemistry, 99:66-72.
[8]	Hu W, Fagundez S, Katin-Grazzini L, Li Y J, Li W, Chen Y N, Wang X M, Deng Z N, Xie S X, McAvoy R J, Li Y. 2017. Endogenous auxin and its manipulation influence in vitro shoot organogenesis of citrus epicotyl explants. Horticulture Research, 4(1):471-483.
[9]	Jia Jin-sheng, Si Long-ting, Han Gui-chao, Yang Hao-ning. 2008. Regeneration in vitro and its influential factors of cucumber(Cucumis sativus L.). Journal of Henan Agricultural Sciences,(6):99-102. (in Chinese)
	贾金生, 司龙亭, 韩贵超, 杨皓宁. 2008. 不同基因型黄瓜离体再生及其影响因素的研究. 河南农业科学,(6):99-102.
[10]	Kim K H, Park J S, Lee B M. 2011. Regeneration potential of immature embryos during seed development in spring and winter wheat genotypes. Korean Journal of Crop Science, 56(3):279-283. doi: 10.7740/kjcs.2011.56.3.279 URL
[11]	Li J, Wu Z, Cui L, Zhang T, Guo Q, Xu J, Jia L, Lou Q, Huang S, Li Z, Chen J F. 2014. Transcriptome comparison of global distinctive features between pollination and parthenocarpic fruit set reveals transcriptional phytohormone cross-talk in cucumber(Cucumis sativus L.). Plant & Cell Physiology, 55(7):1325-1342.
[12]	Liu Chao, Han Li-hong, Chu Hong-long, Dai Dong-qin, Wang Hai-bo, Tang Li-zhou. 2019. Identification and expression analysis of abscisic acid biosynthesis related enzyme genes from pepper. Molecular Plant Breeding, 17(15):4908-4914. (in Chinese)
	刘潮, 韩利红, 褚洪龙, 代冬琴, 王海波, 唐利洲. 2019. 辣椒脱落酸合成相关酶基因的鉴定与表达分析. 分子植物育种, 17(15):4908-4914.
[13]	Liu Jia-fei, Zhang Lu, Meng Yong-jiao, Duan Li-li, Luo Yu-wei, Li Ji, Chen Jin-feng. 2019. Study on cucumber parthenocarpy induced by over-expressing of calmodulin-like protein gene CML25-like. Journal of Nanjing Agricultural University, 42(3):421-429. (in Chinese)
	刘嘉斐, 张璐, 孟永娇, 段莉莉, 罗雨薇, 李季, 陈劲枫. 2019. 过表达钙调素类似蛋白基因CML25-like诱导黄瓜单性结实的研究. 南京农业大学学报, 42(3):421-429.
[14]	Liu Xingwang, Zhai Xuling, Zhang Yaqi, Yin Shuai, Feng Zhongxuan, Ren Huazhong. 2020. A review on genetic and molecular biology of fruit morphogenesis in cucumber. Acta Horticulturae Sinica, 47(9):1793-1809. (in Chinese)
	刘兴旺, 翟许玲, 张亚琦, 尹帅, 冯钟萱, 任华中. 2020. 黄瓜果实形态建成的遗传及分子基础研究进展. 园艺学报, 47(9):1793-1809.
[15]	Liu Yu-xia, Ye De-you, Ou Qiao-ming, Li Min-quan. 2014. Regeneration system establishment from cotyledons in vitro of Cucumis sativus L. Northern Horticulture,(19):113-117. (in Chinese)
	刘玉霞, 叶德友, 欧巧明, 李敏权. 2014. 黄瓜离体子叶再生体系的建立. 北方园艺,(19):113-117.
[16]	Meng Q, Feng H. 2014. Improvement of a high frequency regeneration system for in vitro cultured cotyledons of Chinese cabbage(Brassica rapa L. ssp. pekinensis). Research Journal of Biotechnology, 9(7):85-92.
[17]	Mostafa H, Wang H P, Song J P, Li X X. 2020. Effects of genotypes and explants on garlic callus production and endogenous hormones. Scientific Reports, 10(8):175-183. doi: 10.1038/s41598-019-56899-6 URL
[18]	Park O S, Bae S H, Kim S G, Seo P J. 2019. JA-pretreated hypocotyl explants potentiate de novo shoot regeneration in Arabidopsis. Plant Signaling & Behavior, 14(8):e1618180.
[19]	Pfaffl M W. 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nuclc Acids Research, 9(9):2002-2007.
[20]	Qi X H, Zhu Y M, Li S F, Zhou H X, Xu X W, Xu Q, Chen X H. 2020. Identification of genes related to mesocarp development in cucumber. Horticultural Plant Journal, 6(5):293-300. doi: 10.1016/j.hpj.2020.08.001 URL
[21]	Ren Gui-jie, Dong He-zhong, Chen Yong-zhe, Zhuang Yun-long, Shao Feng-zhi, Liu Zhi-fang. 2002. Studies on endogenous hormone changes in the stem terminal of Gossypium hirsutum during flower bud differentiation. Acta Botanica Boreali-Occidentalia Sinica, 22(2):113-118. (in Chinese)
	任桂杰, 董合忠, 陈永喆, 庄云龙, 邵凤之, 刘志方. 2002. 棉花花芽分化时期茎尖内源激素的变化. 西北植物学报, 22(2):113-118.
[22]	Su Y H, Zhang X S. 2014. The hormonal control of regeneration in plants. Current Topics in Developmental Biology, 108:35-69.
[23]	Takahashi H, Kanayama Y, Zheng M S, Kusano T, Hase S, Ikegami M, Shah J. 2004. Antagonistic interactions between the SA and JA signaling pathways in Arabidopsis modulate expression of defense genes and gene-for-gene resistance to cucumber mosaic virus. Plant & Cell Physiology, 45(6):803-809.
[24]	Tang Y, Liu J, Li X M, Liu B, Li H X. 2011. The influence of endogenous hormones on the formation of buds from stems of bitter melon (Momordca charanta L.). African Journal of Biotechnology, 10(31):5856-5860.
[25]	Tang Y, Liu J, Liu B, Li X M, Li H X. 2010. Endogenous hormone concentrations in explants and calluses of bitter melon(Momordica charantia L.). Interciencia, 35(9):680-683.
[26]	Wang X, Yamada T, Kong F J, Abe Y, Hoshino Y, Sato H, Takamizo T, Kanazawa A, Yamada T. 2011. Establishment of an efficient in vitro culture and particle bombardment-mediated transformation systems in Miscanthus sinensis Anderss.,a potential bioenergy crop. Global Change Biology Bioenergy, 3(4):322-332. doi: 10.1111/gcbb.2011.3.issue-4 URL
[27]	Wang Y, Zhou Q, Zhu G T, Wang S H, Ma Y S, Miao H, Zhang S P, Huang S W, Zhang Z H, Gu X F. 2018. Genetic analysis and identification of a candidate gene associated with in vitro regeneration ability of cucumber. Theoretical and Applied Genetics, 131:2663-2675. doi: 10.1007/s00122-018-3182-7 pmid: 30244395
[28]	Xie Qi-xin, Huang Mei-lian, Wu Xiao-ping, Zhuang Dong-hong. 2008. Plant regeneration from leaves of date plum(Diospyros lotus L.). Scientia Agricultura Sinica,(2):607-612. (in Chinese)
	谢启鑫, 黄美连, 吴晓萍, 庄东红. 2008. 君迁子叶片培养再生植株的研究. 中国农业科学,(2):607-612.
[29]	Zhang Ruo-wei, Gu Xing-fang, Wang Ye, Zhang Sheng-ping, Zhang Bao-xi. 2010. Regeneration system establishment from cotyledons in different cucumber(Cucumis sativus L.)genotypes. Acta Agriculturae Boreali-Sinica, 25(S1):50-54. (in Chinese)
	张若纬, 顾兴芳, 王烨, 张圣平, 张宝玺. 2010. 不同黄瓜基因型子叶再生体系的建立. 华北农学报, 25(S1):50-54.

基因描述 Gene description	基因 Gene	引物序列（5′-3′） Primer sequence
内参基因 Reference gene	Actin	TTCTGGTGATGGTGTGAGTC/GGCAGTGGTGGTGAACATG
生长素相关 Auxin-related	PYROX	AGCCCTGTCGCAAATTAGAT/ CAACTTGGCACATTGCTTCT
	AUX1-1	GAAATAATGGACGCGATGTG/GGAGTGGGTGAGAAGTTGGT
	AUX1-2	GTGAGGCTTCCGGTTGTTAT/ TAACAAGAAGGGCACCAACA
细胞分裂素相关 Cytokinin-related	CYP735A	GCTGAAGGAGATGAATGGGT/ ATTAAGCTTGCCATTGCCTT
	CRE1-1	AGCCATGCTTTCTTCCTGAT/ GTTAACGTTGCTGTGCCATC
	CRE1-2	CGCCTTCAACGATTACTTCA/TGGCTTGAGAAATTCCCATT
赤霉素相关 Gibberellin-related	KAUOX	AGGCCAAGGAAGAACAAGAA/ACTTAGGCCAAGCGTCTCAT
	DEL	AAGTGGAGGAGATGTTTGGG/GATCGTCGTGAATCCTGATG
	GIDW1	GCACCATAGCTCGAAGTGAA/TAGCCTCTTGTACCCTCCCA
脱落酸相关 Abscisic acid-related	NCED1	TCACTCAAACCCAACGCT/ACCAACCCGAAAACTCCT
	NCED2	AAGCATTCCCACACCACA/CATTCTCAACCGCATCAAG
	NCED 6	CTCCCGCTGACACATTCT/CCTCGTCTCGTTCGTTGA
茉莉酸相关 Jasmonic acid-related	COI1	GAACGGGACAGTAGAAGAG/TGGTGGTGTAGCAAAGAG
	PDF1	GCTTTTCCTTTTGGCTCTCA/ATCGCAATGCTTCGTGTTCC
	PDF3	CGAGCACGGTGTTTCAGGGA/CGCATTTATCAGCCGAGCAA

基因描述 Gene description	基因 Gene	引物序列（5′-3′） Primer sequence
内参基因 Reference gene	Actin	TTCTGGTGATGGTGTGAGTC/GGCAGTGGTGGTGAACATG
生长素相关 Auxin-related	PYROX	AGCCCTGTCGCAAATTAGAT/ CAACTTGGCACATTGCTTCT
	AUX1-1	GAAATAATGGACGCGATGTG/GGAGTGGGTGAGAAGTTGGT
	AUX1-2	GTGAGGCTTCCGGTTGTTAT/ TAACAAGAAGGGCACCAACA
细胞分裂素相关 Cytokinin-related	CYP735A	GCTGAAGGAGATGAATGGGT/ ATTAAGCTTGCCATTGCCTT
	CRE1-1	AGCCATGCTTTCTTCCTGAT/ GTTAACGTTGCTGTGCCATC
	CRE1-2	CGCCTTCAACGATTACTTCA/TGGCTTGAGAAATTCCCATT
赤霉素相关 Gibberellin-related	KAUOX	AGGCCAAGGAAGAACAAGAA/ACTTAGGCCAAGCGTCTCAT
	DEL	AAGTGGAGGAGATGTTTGGG/GATCGTCGTGAATCCTGATG
	GIDW1	GCACCATAGCTCGAAGTGAA/TAGCCTCTTGTACCCTCCCA
脱落酸相关 Abscisic acid-related	NCED1	TCACTCAAACCCAACGCT/ACCAACCCGAAAACTCCT
	NCED2	AAGCATTCCCACACCACA/CATTCTCAACCGCATCAAG
	NCED 6	CTCCCGCTGACACATTCT/CCTCGTCTCGTTCGTTGA
茉莉酸相关 Jasmonic acid-related	COI1	GAACGGGACAGTAGAAGAG/TGGTGGTGTAGCAAAGAG
	PDF1	GCTTTTCCTTTTGGCTCTCA/ATCGCAATGCTTCGTGTTCC
	PDF3	CGAGCACGGTGTTTCAGGGA/CGCATTTATCAGCCGAGCAA

基因型 Genotype	IAA（μg · g^-1）			CTK（μg · g^-1）			ABA（μg · g^-1）
基因型 Genotype	0 d		14 d	0 d		14 d	0 d		14 d
C1	173.95 ± 1.54 c	↘	169.60 ± 1.54 c	532.96 ± 2.83 a	↘	516.24 ± 7.54 b	2 167.08 ± 19.34 d	↗	2 245.26 ± 17.73 b
C2	194.82 ± 1.13 b	↘	158.90 ± 3.15 d	515.73 ± 10.73 b	↗	579.21 ± 10.38 a	1 990.62 ± 29.21 e	↗	2 146.98 ± 20.47 c
C3	215.63 ± 3.59 a	↘	138.69 ± 2.47 e	326.10 ± 6.11 f	↗	441.15 ± 9.45 c	1 930.31 ± 24.16 f	↘	1 447.84 ± 26.80 f
C4	150.22 ± 2.89 d	↘	124.02 ± 4.73 f	324.91 ± 7.17 f	-	325.31 ± 4.51 e	1 970.52 ± 33.51 ef	↗	2 086.67 ± 33.73 d
C5	119.38 ± 3.77 f	↗	176.71 ± 2.30 b	370.93 ± 8.99 e	↘	313.01 ± 7.17 e	2 332.38 ± 13.40 c	↗	2 419.49 ± 30.71 a
C6	169.23 ± 1.81 c	↘	141.24 ± 2.99 e	438.77 ± 10.38 c	↗	501.45 ± 9.62 b	2 339.08 ± 29.21 c	↘	2 151.45 ± 30.71 c
C7	122.38 ± 2.26 f	↗	190.03 ± 5.73 a	364.98 ± 11.48 e	↗	430.84 ± 10.26 c	1 858.83 ± 43.60 g	↗	1 961.58 ± 33.73 e
C8	150.52 ± 2.38 d	↘	144.38 ± 1.58 e	513.75 ± 4.76 b	↘	364.98 ± 10.91 d	2 486.50 ± 24.16 b	↘	2 188.30 ± 14.22 c
C9	129.41 ± 5.46 e	↗	195.42 ± 3.59 a	437.18 ± 12.50 c	↘	371.33 ± 9.98 d	2 167.08 ± 43.08 d	↘	1 396.46 ± 43.60 g
C10	171.02 ± 5.05 c	↘	141.39 ± 3.49 e	402.67 ± 13.90 d	↗	580.80 ± 11.25 a	2 654.03 ± 9.48 a	↘	2 040.88 ± 4.74 d
基因型 Genotype	GA			JA
基因型 Genotype	0 d		14 d	0 d		14 d
C1	55.98 ± 0.76 f	↗	70.99 ± 1.09 c	42.07 ± 0.61 c	↘	39.97 ± 0.38 e
C2	53.39 ± 1.07 g	↗	60.53 ± 0.12 f	30.95 ± 0.48 f	↗	53.46 ± 1.65 a
C3	41.92 ± 1.64 i	↗	83.47 ± 0.54 a	40.54 ± 0.75 d	↘	29.77 ± 0.97 g
C4	86.78 ± 0.54 a	↘	65.72 ± 0.98 e	28.44 ± 0.32 gh	↗	48.16 ± 0.55 c
C5	83.18 ± 0.57 b	↘	67.96 ± 0.12 d	51.17 ± 1.18 a	↘	48.20 ± 0.75 c
C6	84.33 ± 1.23 b	↘	61.32 ± 1.15 f	29.37 ± 0.86 g	↗	50.35 ± 0.64 b
C7	60.38 ± 0.33 e	↘	46.10 ± 0.12 g	27.76 ± 0.66 h	↗	45.84 ± 0.70 d
C8	77.98 ± 0.94 c	↘	74.52 ± 0.92 b	48.02 ± 0.94 b	↘	29.12 ± 0.43 g
C9	46.75 ± 1.37 h	↗	70.91 ± 0.70 c	33.42 ± 0.65 e	-	34.49 ± 1.57 f
C10	74.30 ± 0.61 d	-	74.63 ± 0.46 b	50.92 ± 1.20 a	↘	35.10 ± 0.81 f

基因型 Genotype	IAA（μg · g^-1）			CTK（μg · g^-1）			ABA（μg · g^-1）
基因型 Genotype	0 d		14 d	0 d		14 d	0 d		14 d
C1	173.95 ± 1.54 c	↘	169.60 ± 1.54 c	532.96 ± 2.83 a	↘	516.24 ± 7.54 b	2 167.08 ± 19.34 d	↗	2 245.26 ± 17.73 b
C2	194.82 ± 1.13 b	↘	158.90 ± 3.15 d	515.73 ± 10.73 b	↗	579.21 ± 10.38 a	1 990.62 ± 29.21 e	↗	2 146.98 ± 20.47 c
C3	215.63 ± 3.59 a	↘	138.69 ± 2.47 e	326.10 ± 6.11 f	↗	441.15 ± 9.45 c	1 930.31 ± 24.16 f	↘	1 447.84 ± 26.80 f
C4	150.22 ± 2.89 d	↘	124.02 ± 4.73 f	324.91 ± 7.17 f	-	325.31 ± 4.51 e	1 970.52 ± 33.51 ef	↗	2 086.67 ± 33.73 d
C5	119.38 ± 3.77 f	↗	176.71 ± 2.30 b	370.93 ± 8.99 e	↘	313.01 ± 7.17 e	2 332.38 ± 13.40 c	↗	2 419.49 ± 30.71 a
C6	169.23 ± 1.81 c	↘	141.24 ± 2.99 e	438.77 ± 10.38 c	↗	501.45 ± 9.62 b	2 339.08 ± 29.21 c	↘	2 151.45 ± 30.71 c
C7	122.38 ± 2.26 f	↗	190.03 ± 5.73 a	364.98 ± 11.48 e	↗	430.84 ± 10.26 c	1 858.83 ± 43.60 g	↗	1 961.58 ± 33.73 e
C8	150.52 ± 2.38 d	↘	144.38 ± 1.58 e	513.75 ± 4.76 b	↘	364.98 ± 10.91 d	2 486.50 ± 24.16 b	↘	2 188.30 ± 14.22 c
C9	129.41 ± 5.46 e	↗	195.42 ± 3.59 a	437.18 ± 12.50 c	↘	371.33 ± 9.98 d	2 167.08 ± 43.08 d	↘	1 396.46 ± 43.60 g
C10	171.02 ± 5.05 c	↘	141.39 ± 3.49 e	402.67 ± 13.90 d	↗	580.80 ± 11.25 a	2 654.03 ± 9.48 a	↘	2 040.88 ± 4.74 d
基因型 Genotype	GA			JA
基因型 Genotype	0 d		14 d	0 d		14 d
C1	55.98 ± 0.76 f	↗	70.99 ± 1.09 c	42.07 ± 0.61 c	↘	39.97 ± 0.38 e
C2	53.39 ± 1.07 g	↗	60.53 ± 0.12 f	30.95 ± 0.48 f	↗	53.46 ± 1.65 a
C3	41.92 ± 1.64 i	↗	83.47 ± 0.54 a	40.54 ± 0.75 d	↘	29.77 ± 0.97 g
C4	86.78 ± 0.54 a	↘	65.72 ± 0.98 e	28.44 ± 0.32 gh	↗	48.16 ± 0.55 c
C5	83.18 ± 0.57 b	↘	67.96 ± 0.12 d	51.17 ± 1.18 a	↘	48.20 ± 0.75 c
C6	84.33 ± 1.23 b	↘	61.32 ± 1.15 f	29.37 ± 0.86 g	↗	50.35 ± 0.64 b
C7	60.38 ± 0.33 e	↘	46.10 ± 0.12 g	27.76 ± 0.66 h	↗	45.84 ± 0.70 d
C8	77.98 ± 0.94 c	↘	74.52 ± 0.92 b	48.02 ± 0.94 b	↘	29.12 ± 0.43 g
C9	46.75 ± 1.37 h	↗	70.91 ± 0.70 c	33.42 ± 0.65 e	-	34.49 ± 1.57 f
C10	74.30 ± 0.61 d	-	74.63 ± 0.46 b	50.92 ± 1.20 a	↘	35.10 ± 0.81 f

激素类型 Hormone types	分化率 Differentiation rate		平均分化芽数 Average bud number
激素类型 Hormone types	0 d	14 d	0 d	14 d
IAA	0.1076	0.0086	0.0012	-0.0164
CTK	0.4544	0.6110	0.1863	-0.0184
ABA	0.0266	0.1172	0.0043	0.0001
GA	-0.0392	-0.0074	-0.0083	0.1070
JA	0.0057	0.0307	0.0214	-0.1026