十一个桑黄菌株的形态学观察与分子鉴定

doi:10.3969/j.issn.1004-1524.20230417

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (4): 800-810.DOI: 10.3969/j.issn.1004-1524.20230417

十一个桑黄菌株的形态学观察与分子鉴定

陆春霞¹^,²^,³(), 刘开莉¹^,²^,³, 梁贵秋¹^,²^,³, 肖潇¹^,²^,³, 韦伟¹^,²^,³, 徐雯雯¹^,²^,³, 陈菁¹^,²^,³, 莫炳巧¹^,²^,³, 李小群¹^,²^,³, 黎尔纳⁴^,⁵^,⁶, 黄旭华¹^,²^,³, 李安华¹^,²^,³, 韦师妮¹^,²^,³

1.广西壮族自治区蚕业技术推广站,广西南宁 530007
2.广西壮族自治区蚕业科学研究院,广西南宁 530007
3.广西蚕桑遗传改良与高效种养重点实验室,广西南宁 530007
4.广东省农业科学院蚕业与农产品加工研究所,广东广州 510610
5.农业农村部功能食品重点实验室,广东广州 510610
6.广东省农产品加工重点实验室,广东广州 510610

收稿日期:2023-03-31 出版日期:2024-04-25 发布日期:2024-04-29
作者简介:陆春霞(1976—),女,广西宁明人,硕士,正高级农艺师,主要从事蚕桑资源综合利用研究。E-mail:75727907@qq.com
基金资助:
广西重点研发计划项目(桂科AB21196040)

Morphological observation and molecular identification of 11 sanghuang strains

LU Chunxia¹^,²^,³(), LIU Kaili¹^,²^,³, LIANG Guiqiu¹^,²^,³, XIAO Xiao¹^,²^,³, WEI Wei¹^,²^,³, XU Wenwen¹^,²^,³, CHEN Jing¹^,²^,³, MO Bingqiao¹^,²^,³, LI Xiaoqun¹^,²^,³, LI Er’na⁴^,⁵^,⁶, HUANG Xuhua¹^,²^,³, LI Anhua¹^,²^,³, WEI Shini¹^,²^,³

1. Guangxi Zhuang Autonomous Region Sericultural Technology Promotion Station, Nanning 530007, China
2. Guangxi Research Academy of Sericultural Science, Nanning 530007, China
3. Guangxi Key Laboratory of Sericultural Genetic Improvement and Efficient Breeding, Nanning 530007, China
4. Sericultural & Agri-Food Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510610, China
5. Key Laboratory of Functional Foods, Ministry of Agriculture and Rural Affairs, Guangzhou 510610, China
6. Guangdong Key Laboratory of Agricultural Products Processing, Guangzhou 510610, China

Received:2023-03-31 Online:2024-04-25 Published:2024-04-29

摘要/Abstract

摘要：

为明确桑黄菌株的种属,对采集自国内不同地区的11个桑黄类似菌株开展分子鉴定和形态学特性观察,主要采用核糖体基因rDNA内转录间隔区(ITS)对桑黄菌株进行基因鉴定和遗传性状分析,运用Mega7.0软件的neighbor-joining(NJ)法构建基于桑黄rDNA ITS序列的系统发育进化树,结合桑黄菌株的形态学观察(菌丝生长情况、菌丝的显微结构、孢子形态特征、子实体电镜扫描结构)进行分析鉴定。结果表明:11个桑黄菌株分为两大类群,其中菌株SH-1为一类,属于纤孔菌属(Inonotus);其他菌株(SH-2、SH-3、SH-4、SH-5、SH-6、SH-7、SH-8、SH-9、SH-10、SH-11)属于另一类,为桑黄孔菌属(Sanghuangporus),菌株SH-3和SH-6属于桑树桑黄(S. sanghuang),其余8个菌株属于暴马桑黄(S. baumii)。通过rDNA ITS序列分析技术结合桑黄菌丝形态特征、孢子形态特征和子实体显微结构特征分析,能明确桑黄的种属与遗传性状,可为今后桑黄品种选育与人工规模化生产提供理论依据。

关键词: 桑黄, 纤孔菌属, 桑黄孔菌属, 桑树桑黄, 暴马桑黄, 分子鉴定

Abstract:

In order to clarify the genus of 11 similar sanghuang strains collected from different regions in China, molecular identification and morphological characterization observation were carried out. The ribosomal gene rDNA intertranscriptional spacer (ITS) technique was used to identify the genes and analyze the genetic traits of 11 sanghuang strains. Mega 7.0’s neighbor-joining (NJ) method was used to construct phylogenetic tree based on rDNA ITS sequence of 11 sanghuang strains, and the morphological observation (mycelium growth, mycelium microstructure, spore morphological characteristics, fruite body scanning structure) was analyzed and identified. The results showed that the 11 sanghuang strains were divided into two major groups, of which the strain SH-1 classified into one group, belonging to the genus Inonotus; the others strains (SH-2, SH-3, SH-4, SH-5, SH-6, SH-7, SH-8, SH-9, SH-10 and SH-11) belonged to Sanghuangporus, the strains SH-3 and SH-6 belonged to S. sanghuang, and the other 8 strains belonged to S. baumii. This method is a combination of rDNA ITS sequence analysis technology and morphological characteristics of mycelia, spore morphological characteristics and fruit-body microscopic structure characteristics of sanghuang, which can clarify the species and genetic characteristics of sanghuang, and can be used as an important theoretical basis for the artificial large-scale production of sanghuang in the future.

Key words: sanghuang, Inonotus, Sanghuangporus, Sanghuangporus sanghuang, Sanghuangporus baumii, molecular identification

中图分类号:

S567.3

陆春霞, 刘开莉, 梁贵秋, 肖潇, 韦伟, 徐雯雯, 陈菁, 莫炳巧, 李小群, 黎尔纳, 黄旭华, 李安华, 韦师妮. 十一个桑黄菌株的形态学观察与分子鉴定[J]. 浙江农业学报, 2024, 36(4): 800-810.

LU Chunxia, LIU Kaili, LIANG Guiqiu, XIAO Xiao, WEI Wei, XU Wenwen, CHEN Jing, MO Bingqiao, LI Xiaoqun, LI Er’na, HUANG Xuhua, LI Anhua, WEI Shini. Morphological observation and molecular identification of 11 sanghuang strains[J]. Acta Agriculturae Zhejiangensis, 2024, 36(4): 800-810.

图/表 10

表1 十一个桑黄菌株来源与编号

Table 1 Number and source of 11 sanghuang strains

菌株编号 Strain number	菌株来源 Source of strain
SH-1	湖南,桑属Hunan, Morus
SH-2	新疆,丁香属Xinjiang, Syringa
SH-3	浙江,桑属Zhejiang, Moru
SH-4	浙江,丁香属Zhejiang, Syringa
SH-5	浙江,丁香属Zhejiang, Syringa
SH-6	山东,桑属Shandong, Moru
SH-7	浙江,丁香属Zhejiang, Syringa
SH-8	吉林,丁香属Jilin, Syringa
SH-9	浙江,丁香属Zhejiang, Syringa
SH-10	湖南,丁香属Hunan, Syringa
SH-11	新疆,丁香属Xinjiang, Syringa

图1 桑黄菌株的菌落形态特征 A,菌丝生长5 d的形态;B,菌丝生长15 d的形态。

Fig.1 The colony morphological characteristics of sanghuang strains A, Morphology of mycelium growth for 5 d; B, Morphology of mycelium growth for 15 d.

图2 桑黄菌丝显微形态特征

Fig.2 Microscopic morphological characteristics of sanghuang strains mycelia

图3 桑黄子实体

Fig.3 Fruiting body of sanghuang strains

图4 桑黄子实体横纵切片的扫描电镜图

Fig.4 Scanning electron microscope images of transverse and longitudinal sections of fruiting body of sanghuang strains

图5 桑黄孢子显微结构图

Fig.5 Micrograph of the spore of sanghuang strains

图6 桑黄菌丝基因组DNA的凝胶电泳图

Fig.6 Agarose gel electrophoresis of genomic DNA of 11 sanghuang strains M, DNA marker.

图7 桑黄菌丝rDNA ITS序列扩增电泳结果

Fig.7 Electrophoresis analysis of amplified product of sanghuang strains rDNA ITS sequences M, DNA marker.

图8 基于桑黄真菌rDNA ITS序列的系统发育进化树

Fig.8 Phylogenetic tree based on rDNA ITS sequences of sanghuang strains

表2 各菌种序列间的遗传距离

Table 2 Genetic distance among sanghuang strains

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十一个桑黄菌株的形态学观察与分子鉴定

Morphological observation and molecular identification of 11 sanghuang strains

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