[1]高 晗,王毅敏,李 薇,等.杜鹃兰体胚发生细胞学观察和生理生化差异[J].南京师大学报(自然科学版),2026,49(02):59-66.[doi:10.3969/j.issn.1001-4616.2026.02.006]
　Gao Han,Wang Yimin,Li Wei,et al.Cytological Observation and Physiological and Biochemical Difference of Embryogenesis in Callus of Cremastra appendiculata[J].Journal of Nanjing Normal University(Natural Science Edition),2026,49(02):59-66.[doi:10.3969/j.issn.1001-4616.2026.02.006]
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杜鹃兰体胚发生细胞学观察和生理生化差异()

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
49
期数:
2026年02期
页码:
59-66
栏目:
生物学
出版日期:
2026-04-10

文章信息/Info

Title:
Cytological Observation and Physiological and Biochemical Difference of Embryogenesis in Callus of Cremastra appendiculata
文章编号:
1001-4616(2026)02-0059-08
作者:
高 晗1; 王毅敏1; 李 薇1; 梁宏伟2; 杨艾菊3; 高本旺1
1.三峡植物园,湖北 宜昌 443100
2.三峡大学生物与制药学院,湖北 宜昌 443002
3.宜昌市夷陵区樟村坪林场,湖北 宜昌 443100
Author(s):
Gao Han1; Wang Yimin1; Li Wei1; Liang Hongwei2; Yang Aiju3; Gao Benwang1
1.Three Gorges Botanical Garden,Yichang 443100,China
2.College of Biological and Pharmaceutical Sciences of China Three Gorges University,Yichang 443002,China
3.Zhangcunping Forest Farm in Yiling District,Yichang 443100,China
关键词:
杜鹃兰; 体胚发生; 细胞学观察; 生理生化
Keywords:
C.appendiculata;  somatic embryogenesis;  cytological observation;  physiology and biochemistry
分类号:
S682.31
DOI:
10.3969/j.issn.1001-4616.2026.02.006
文献标志码:
A
摘要:
本研究以建立稳定的杜鹃兰(Cremastra appendiculata)体胚发生和再生体系,促进珍稀药用植物杜鹃兰种质资源保存和规模化繁殖,为其遗传转化提供依据. 以杜鹃兰未成熟幼胚为外植体,诱导杜鹃兰愈伤组织,研究不同浓度及配比(IBA与6-BA组合)的植物生长调节剂对愈伤组织增殖的影响; 对不同形态的愈伤组织进行了形态学和生理生化差异分析(可溶性糖、可溶性蛋白及抗氧化酶活性检测); 利用体视显微镜观察和石蜡切片技术详细研究了体细胞胚的起源及发育过程. 结果显示,杜鹃兰种子可通过1/2 MS培养基诱导出淡黄色愈伤组织,愈伤组织增殖最适培养基为MS+(0.5 mg/L、1 mg/L)IBA+(2 mg/L、3 mg/L)6-BA,增殖形成的胚性愈伤组织可在添加0.2 mg/L IBA的1/2 MS培养基上诱导再生植株,通过体视显微镜及组织切片观察,杜鹃兰细胞胚经过球形胚、梨形胚、凹沟胚和子叶胚阶段,与合子胚发育过程类似. 不同形态愈伤组织可溶性糖、可溶性蛋白及抗氧化酶活性具有显著差异,其中Ⅰ型愈伤组织胚状体发生潜力最大,Ⅰ型、Ⅲ型愈伤组织抗逆性最强. 本研究成功建立了杜鹃兰的体细胞胚胎发生与植株再生体系,确定了关键的培养基配方(诱导、增殖和再生). 结果揭示了不同形态愈伤组织的生理生化特性及其与胚状体发生潜力和抗逆性的关联,并明确了杜鹃兰体细胞胚与合子胚发育过程的相似性.
Abstract:
The study was to establish a stable somatic embryogenesis and regeneration system for Cremastra appendiculata, promote the conservation of germplasm resources and large-scale propagation of this rare medicinal plant, and provide a basis for its genetic transformation. Immature embryos of C.appendiculata were used as explants to induce callus formation. The effects of plant growth regulators at various concentrations and ratios(combinations of IBA and 6-BA)on callus proliferation were systematically investigated. Morphological and physiological-biochemical differences among various callus types were analyzed, including assays of soluble sugars, soluble proteins, and antioxidant enzyme activities. Furthermore, stereomicroscopic observation and paraffin sectioning techniques were employed to examine the origin and developmental process of somatic embryos in detail. Seeds of C.appendiculata could induce pale-yellow callus on 1/2 MS medium. The optimal medium for callus proliferation was MS supplemented with IBA(0.5 mg/L, 1 mg/L)combined with 6-BA(2 mg/L, 3 mg/L). Embryogenic callus formed under these conditions could regenerate plants on 1/2 MS medium containing 0.2 mg/L IBA. Observations via stereomicroscopy and tissue sections revealed that C.appendiculata somatic embryos developed through globular, pear-shaped, cotyledonary and mature embryo stages, similar to the development of zygotic embryos. Significant differences in soluble sugar, soluble protein content and antioxidant enzyme activities were observed among callus types. Type I callus exhibited the highest embryoid potential, whereas Types I and III callus showed the strongest stress resistance. This study successfully established a somatic embryogenesis and plant regeneration system for C.appendiculata and identified key medium formulations for induction, proliferation and regeneration. The results reveal the physiological and biochemical characteristics of different callus types and their correlation with embryoid potential and stress tolerance, and confirm the similarity between the developmental processes of somatic and zygotic embryos in C.appendiculata.

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备注/Memo

备注/Memo:
收稿日期:2025-10-20.
基金项目:2024年湖北省林业生态文明建设资金项目(42000021816T300000130)、湖北省重点实验室开放课题项目(SNJGKL202103)、湖北省科技厅中央引导地方科技发展项目(2022BGE265).
通讯作者:高本旺,高级工程师,研究方向:植物引种驯化及应用. E-mail:953146673@qq.com
更新日期/Last Update: 2026-04-10