Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (8): 1891-1905.doi: 10.16420/j.issn.0513-353x.2023-0621
• Cultivation·Physiology & Biochemistry • Previous Articles Next Articles
WANG Jianzhao, GAO Yuan, DAO Mei, ZHANG Hongye, YANG Ziyun, CHEN Longqing, WU Tian*()
Received:
2023-10-24
Revised:
2024-04-23
Online:
2024-08-25
Published:
2024-08-22
Contact:
WU Tian
WANG Jianzhao, GAO Yuan, DAO Mei, ZHANG Hongye, YANG Ziyun, CHEN Longqing, WU Tian. Establishment of Regeneration in Vitro System of Camellia japonica and Relationship Between Endogenous Hormones and Adventitious Bud Differentiation[J]. Acta Horticulturae Sinica, 2024, 51(8): 1891-1905.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2023-0621
培养基编号 Medium number | 培养基配方/(mg · L-1) Medium component | 诱导率/% Induction percentage | ||||
---|---|---|---|---|---|---|
6-BA | 2,4-D | NAA | IBA | KT | ||
YS1 | 2.0 | 1.5 | 0.05 | 0 | 0 | 42.68 |
YS2 | 2.5 | 2.0 | 0.50 | 0 | 0 | 35.20 |
YS3 | 5.0 | 0.1 | 0 | 0 | 0 | 79.19 |
YS4 | 2.0 | 0.5 | 0 | 0 | 0 | 41.54 |
YS5 | 0.4 | 2.0 | 0 | 0 | 0 | 43.60 |
YS6 | 0.5 | 0 | 2.00 | 0.1 | 1.0 | 66.57 |
Table 1 Callus induction media from Camellia japonica leaves
培养基编号 Medium number | 培养基配方/(mg · L-1) Medium component | 诱导率/% Induction percentage | ||||
---|---|---|---|---|---|---|
6-BA | 2,4-D | NAA | IBA | KT | ||
YS1 | 2.0 | 1.5 | 0.05 | 0 | 0 | 42.68 |
YS2 | 2.5 | 2.0 | 0.50 | 0 | 0 | 35.20 |
YS3 | 5.0 | 0.1 | 0 | 0 | 0 | 79.19 |
YS4 | 2.0 | 0.5 | 0 | 0 | 0 | 41.54 |
YS5 | 0.4 | 2.0 | 0 | 0 | 0 | 43.60 |
YS6 | 0.5 | 0 | 2.00 | 0.1 | 1.0 | 66.57 |
培养基编号 Medium number | 培养基配方/(mg · L-1)Medium component | 生根率/% Induction percentage of rooting | |
---|---|---|---|
IAA | IBA | ||
Sg1 | 0.5 | 0 | 0 |
Sg2 | 0.5 | 0 | 33.33 |
Sg3 | 0 | 0.5 | 50.00 |
Table 2 Induction of adventitious root media for Camellia japonica
培养基编号 Medium number | 培养基配方/(mg · L-1)Medium component | 生根率/% Induction percentage of rooting | |
---|---|---|---|
IAA | IBA | ||
Sg1 | 0.5 | 0 | 0 |
Sg2 | 0.5 | 0 | 33.33 |
Sg3 | 0 | 0.5 | 50.00 |
Fig. 1 Overall process of in vitro regeneration from Camellia japonica leaves a,b:Camellia japonica callus in YS3 and YS6 media;c:Callus after three times of subculture;d:Differentiated adventitious buds;e:Subculture of adventitious bud;f:Segmentation of adventitious buds;g:Adventitious roots could not be induced on Sg1 rooting media;h,i:Adventitious roots on Sg2 and Sg3 rooting media.
培养基编号 Medium number | 基质配比 Matrix ratio | 成活率/% Rate of survival | |||
---|---|---|---|---|---|
腐殖土 Humus | 珍珠岩 Perlite | 蛭石 Vermiculite | 生土 Raw soil | ||
1 | 1 | 1 | 1 | 3 | 66.7 |
2 | 1 | 1 | 0 | 2 | 66.7 |
3 | 0 | 1 | 0 | 1 | 66.7 |
4 | 0 | 0 | 0 | 1 | 0 |
Table 3 Transplant survival of Camellia japonica grouped seedlings in different substrates
培养基编号 Medium number | 基质配比 Matrix ratio | 成活率/% Rate of survival | |||
---|---|---|---|---|---|
腐殖土 Humus | 珍珠岩 Perlite | 蛭石 Vermiculite | 生土 Raw soil | ||
1 | 1 | 1 | 1 | 3 | 66.7 |
2 | 1 | 1 | 0 | 2 | 66.7 |
3 | 0 | 1 | 0 | 1 | 66.7 |
4 | 0 | 0 | 0 | 1 | 0 |
Fig. 2 Endogenous hormone clustering heat map of callus content difference between Camellia japonica with adventitious bud differentiation(with bud)and without adventitious bud differentiation(without bud) Red represents high content,green represents low content. The numbers 1,2 and 3 indicate three biological repeats.
Fig. 3 Differential endogenous hormone KEGG enrichment map of callus content in Camellia japonica with adventitious bud differentiation(with bud)and without adventitious bud differentiation(without bud) The red indicates the more significant enrichment. The size of the dots represents the number of differentiated metabolites enriched.
Fig. 4 Auxin hormone content in callus with adventitious bud differentiation(with bud)and without adventitious bud differentiation(without bud)in Camellia japonica IAA-Glu:Indoleacetic acid-glutamic acid;IAA-Val-Me:Indole-3-acetyl-L-valine methyl ester;IPA:3-Indolepopionic acid;ICAld:Indole-3-formaldehyde;IAA-Ala:N-(3-Indolyl acetyl)-L-alanine;TRA:Tryptamine. The bar plot represents the mean of the 3 biological replicates,and the error bar is the standard deviation. * represents the statistical difference in the t-test. * P < 0.05;** P < 0.01;*** P < 0.001;**** P < 0.0001. The same below.
Fig. 5 Cytokinins in callus with adventitious bud differentiation(with bud)and without adventitious bud differentiation(without bud)in Camellia japonica K9G:Kinetin-9-glucoside;DHZROG:Dihydrozeatin-O-glucoside riboside;mT:meta-Topolin;2MeScZR:2-Methylthio-cis-zeatin riboside;pT9G:4-{[(9-β-D-Glucopyranosyl-9H-purin-6-yl)amino]methyl}phenol;BAPR:6-Benzyladenosine;mTR:meta-Topolin riboside;IPR:Isopentenyl adenine nucleoside;oT:6-(2-Hydroxybenzyl amino);iP9G:Isopentenadenine-9-glucoside;tZOG:trans-Zeatin-O-glucoside;DHZ7G:Dihydrozeaxin-7-glycoside;iP7G:Isopentenadenine-7-glucoside;oTR:ortho-Topolin;pTR:para-Topolin riboside;cZROG:cis-Zeatin-O-glucoside riboside;IP:N6-isopentennyladenine;mT9G:meta-Topolin-9-glucoside;BAP9G:N6-Benzyladenine-9-glucoside.
Fig. 6 Jasmonates in callus with adventitious bud differentiation(with bud)and without adventitious bud differentiation(without bud)in Camellia japonica JA-Val:Jasmonate-valine;JA-Phe:Jasmonate-phenylalanine;JA-ILE:Jasmonate-isoleucine;JA:Jasmonate;OPC-4:Oxypentenyl cyclopentane butyric acid.
Fig. 7 Salicylic acids in callus with adventitious bud differentiation(with bud)and without adventitious bud differentiation(without bud)in Camellia japonica SAG:Salicylate 2-O-β-glucoside;SA:Salicylic acid;ST:Strigol.
Fig. 8 Model diagram of regeneration in vitro of Camellia japonica ↑ means increased endogenous hormone levels;↑ means reduced endogenous hormone levels.
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