Selection and Sequence Alignment of PCR Primers for Identifying Zizhi Strain

ZHONG Liyi; CHEN Tiqiang; LIU Xinrui; YIN Zhenghe; YNAG Chi

doi:10.19303/j.issn.1008-0384.2020.07.005

Volume 35 Issue 7

Jul. 2020

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Article Navigation > Fujian Journal of Agricultural Sciences > 2020 > 35(7): 725-730

ZHONG L Y, CHEN T Q, LIU X R, et al. Selection and Sequence Alignment of PCR Primers for Identifying Zizhi Strain [J]. Fujian Journal of Agricultural Sciences，2020，35（7）：725−730 doi: 10.19303/j.issn.1008-0384.2020.07.005

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ZHONG L Y, CHEN T Q, LIU X R, et al. Selection and Sequence Alignment of PCR Primers for Identifying Zizhi Strain [J]. Fujian Journal of Agricultural Sciences，2020，35（7）：725−730 doi: 10.19303/j.issn.1008-0384.2020.07.005

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Selection and Sequence Alignment of PCR Primers for Identifying Zizhi Strain

doi: 10.19303/j.issn.1008-0384.2020.07.005

1.
Fujian Wuping Mushroom Extension Station, Wuping, Fujian 364000, China
2.
Edible & Medicinal Fungi Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350014, China
3.
Mycological Research Center, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China

Received Date: 2020-01-16
Rev Recd Date: 2020-06-03
Publish Date: 2020-07-31

Abstract

Abstract

Objective The molecular markers of the cultivated strain of Ganoderma sinense, Zizhi S2 (aka Wu-Zhi No. 2), recently popularized in Fujian and surrounding provinces were studied to facilitate the authentication of the medicinal fungus. Method Relevant primers of Zizhi S2 showing clear and stable bands and polymorphism were screened using PCR. Phylogenetic tree of UPMGA clustering analysis on the verified authentic cultivars was constructed to determine their relationship by genetic distance as well as antagonistic reaction. Subsequently, sequences of the selected primers were blasted on the genome of Zizhi S2 to validate the methodology. Result There were 2 RAPD-PCR and 3 ISSR-PCR primers found to clearly and stably amplify the specific or polymorphic bands. However, the sites and numbers on the scaffolds of the Zizhi S2 genome that matched the sequences of the 5 primers were not same. Conclusion It was confirmed that three primers(ISSR13, S1326, and S1506)could be effectively used for identification of Zizhi cultivated strains based on sequences alignment with the genome of G. sinense strain Zizhi S2.
- Ganoderma sinense,
- cultivated strain,
- molecular markers,
- PCR primers,
- sequence alignment

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References(21)

References

[1]	李开本, 陈体强, 何修金, 等. 短段木灵芝优质高产菌株“赤芝6号”研究 [J]. 福建省农科院学报, 1996, 11(3):1−6. LI K B, CHEN T Q, HE X J, et al. Study on“Red Reishi No. 6”, a high-yield and fine-quality strain of Ganoderma lucidum for woodlqg cultivation [J]. Fujian Journal of Agricultural Sciences, 1996, 11(3): 1−6.(in Chinese)
[2]	林兴生, 李开本, 陈体强, 等. 28个灵芝菌株栽培特性比较 [J]. 食用菌学报, 2001, 8(3):45−49. doi: 10.3969/j.issn.1005-9873.2001.03.009 LIN X S, LI K B, CHEN T Q, et al. A comparative study on the cultivation characteristics of 28 Ganoderma lucidum strains [J]. Acta Edulis Fungi, 2001, 8(3): 45−49.(in Chinese) doi: 10.3969/j.issn.1005-9873.2001.03.009
[3]	陈体强, 林兴生, 赵健, 等. 福建灵芝科真菌资源及担孢子形态结构数据库研究 [J]. 福建农业学报, 2002, 17(1):40−44. doi: 10.3969/j.issn.1008-0384.2002.01.011 Chen T Q, Lin X S, ZHAO J, et al. Resource of Ganodermataceae in Fujian Province and its morphology and structure database of basidospores [J]. Fujian Journal of Agricultural Sciences, 2002, 17(1): 40−44.(in Chinese) doi: 10.3969/j.issn.1008-0384.2002.01.011
[4]	赵继鼎. 中国灵芝科的分类研究: XI.灵芝亚属灵芝组 [J]. 真菌学报, 1989(1):25−34. ZHAO J D. Studies on the taxonomy of Ganodermataceae in China vi [J]. Mycosystema, 1989(1): 25−34.(in Chinese)
[5]	赵继鼎, 张小青. 中国真菌志(第十八卷)·灵芝科[M]. 北京: 科学出版社, 2000, 70: 86－87&103－106.
[6]	李钦艳, 钟莹莹, 陈逸湘, 等. 我国灵芝种质资源及生产技术研究进展 [J]. 中国食用菌, 2016, 35(1):8−12, 20. LI Q Y, ZHONG Y Y, CHEN Y X, et al. Research advances on the production technology of Ganoderma in China [J]. Edible Fungi of China, 2016, 35(1): 8−12, 20.(in Chinese)
[7]	陈逸湘, 宋斌, 李挺, 等. 紫芝研究进展 [J]. 广东农业科学, 2011, 38(24):36−39. doi: 10.3969/j.issn.1004-874X.2011.24.014 CHEN Y X, SONG B, LI T, et al. Research on advances of Ganoderma sinense [J]. Guangdong Agricultural Sciences, 2011, 38(24): 36−39.(in Chinese) doi: 10.3969/j.issn.1004-874X.2011.24.014
[8]	陈体强, 吴锦忠, 李晔, 等. 福建野生紫芝资源开发利用Ⅱ.“闽紫96”(中国灵芝) [J]. 菌物研究, 2006, 4(4):27−32. doi: 10.3969/j.issn.1672-3538.2006.04.005 CHEN T Q, WU J Z, LI Y, et al. Exploitation and utilization of wild Zhizi resources(sect. phaeonema of Ganoderma) in Fujian (Ⅱ)/. G. sinense “minzi 96” [J]. Journal of Fungal Research, 2006, 4(4): 27−32.(in Chinese) doi: 10.3969/j.issn.1672-3538.2006.04.005
[9]	钟礼义, 陈体强, 邱福平. 不同紫芝菌株特性比较及边材栽培研究初报 [J]. 食用菌, 2005, 27(1): 26－27. ZHONG L Y, CHEN T Q, QIU F P, et al. Primary study on cultivation of Zizhi with broad-leaved sapwood and comparative characteristics of different strains[J]. Edible Fungi, 2005, 27(1): 26－27. (in Chinese)
[10]	吴兴亮, 戴玉成. 中国灵芝图鉴 [M]. 北京: 科学出版社, 2005.
[11]	王新存. 灵芝科系统发育研究 [D]. 北京: 中国科学院大学, 2012. WANG X C. Phylogenetic studies of Ganodermataceae [D]. Beijing: University of Chinese Academy of Sciences, 2012.(in Chinese)
[12]	HAPUARACHCHI K K. High phenotypic plasticity of Ganoderma sinense (ganodermataceae, polyporales) in China [J]. Asian Journal of Mycology, 2019, 2(1): 1−47. doi: 10.5943/ajom/2/1/1
[13]	钟礼义. 紫芝新品种武芝2号区域试验 [J]. 中国食用菌, 2013, 32(6):25−27. ZHONG L Y. The regional test on new varieties of Zizhi (Ganoderma) [J]. Edible Fungi of China, 2013, 32(6): 25−27.(in Chinese)
[14]	应正河. RAPD、SRAP和ISSR标记在香菇种质资源的应用及其SCAR标记的建立 [D]. 福州: 福建农林大学. 2006. YIN Z H. Application of RAPD, SRAP and ISSR markers in germplasm resources of Lentinus edodes and establishment of SCAR markers [D]. Fuzhou: Fujian Agriculture and Forestry University, 2005. (in Chinese)
[15]	唐传红, 张劲松, 陈明杰, 等. 利用拮抗试验和RAPD对灵芝属菌株进行分类研究 [J]. 微生物学通报, 2005, 32(5):72−76. doi: 10.3969/j.issn.0253-2654.2005.05.015 TANG C H, ZHANG J S, CHEN M J, et al. Study on classification of strains of Ganoderma by anatagonistic effect and rapd [J]. Microbiology, 2005, 32(5): 72−76.(in Chinese) doi: 10.3969/j.issn.0253-2654.2005.05.015
[16]	王锦锋, 李晶, I.L.Datti, et al 利用拮抗、ITS和RAPD技术对灵芝属菌株分类的研究 [J]. 西南农业学报, 2017, 30(1):26−33. WANG J F, LI J, DATTI I, et al. Study on classification of 21 strains of Ganoderma by antagonistic effect, ITS and RAPD technology [J]. Southwest China Journal of Agricultural Sciences, 2017, 30(1): 26−33.(in Chinese)
[17]	徐凯, 唐传红, 王天娇, 等. 基于SRAP、ISSR和RAPD分析灵芝G0130菌株单核体多态性 [J]. 工业微生物, 2014, 44(4):39−45. doi: 10.3969/j.issn.1001-6678.2014.04.008 XU K, TANG C H, WANG T J, et al. Polymorphism of monokaryons of Ganoderma lucidum G0130 based on SRAP, ISSR and RAPD analysis [J]. Industrial Microbiology, 2014, 44(4): 39−45.(in Chinese) doi: 10.3969/j.issn.1001-6678.2014.04.008
[18]	LIU D B, GONG J, DAI W K, et al. The genome of Ganoderma lucidum provides insights into triterpenes biosynthesis and wood degradation [J]. PLoS One, 2012, 7(5): e36146. doi: 10.1371/journal.pone.0036146
[19]	CHEN S L, XU J, LIU C, et al. Genome sequence of the model medicinal mushroom Ganoderma lucidum [J]. Nature Communications, 2012, 3: 913. doi: 10.1038/ncomms1923
[20]	ZHU Y J, XU J, SUN C, et al. Chromosome-level genome map provides insights into diverse defense mechanisms in the medicinal fungus Ganoderma sinense [J]. Scientific Reports, 2015, 5: 11087. doi: 10.1038/srep11087
[21]	龚明, 鲍大鹏, 唐传红, 等. 沪农灵芝1号全基因组分析和演化比较 [J]. 食用菌学报, 2016, 23(2):1−5, 19. GONG M, BAO D P, TANG C H, et al. Genome-wide analysis and evolution comparison of Ganoderma lucidum SH [J]. Acta Edulis Fungi, 2016, 23(2): 1−5, 19.(in Chinese)