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| SSR Identification of Transcriptome and Fingerprint Construction in Color-leafed Cinnamomum camphora |
| KONG Lin-Lin1, MENG Xian-Dong2, WANG Jian-Jun3, YU Jin-Jian1, HU Xian-Ge1, LIN Er-Pei1, HUANG Hua-Hong1,* |
1 The State Key Laboratory of Subtropical Silviculture, Zhengjiang Agricultural and Forestry University, Hangzhou 311300, China;
2 Forestry Seed and Seedling Management Station of Zhejiang Province, Hangzhou 311300, China;
3 Ningbo Agricultural Technology Extension Station , Ningbo 315012, China |
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Abstract Color-leafed Cinnamomum camphora is an ornamental plant variety with colored leaves in spring. Although these varieties have broad market prospects, a technology system for distinguishing them effectively has not yet been established, which affects the protection of new variety rights and market circulation. In this study, in order to rapidly identify 3 color-leafed varieties, their fingerprints were constructed using SSR markers derived from the transcriptome of C. camphora. The transcriptome of C. camphora 'Yongjin' was sequenced using the Illumina high-throughput sequencing platform, and a total of 129 175 Unigenes were obtained, with a total length of 92 429 171 bp. The SSR search of the transcriptome sequences by MIcroSAtellite identification tool (MISA) revealed that 28 895 sequences contained 36 430 SSR sites, with an average of 1 SSR per 2 537.2 bp and a frequency of 22.37%. Mononucleotide repeats (accounting for 49.74%) were the most abundant, followed by dinucleotide (29.12%) and trinucleotide (19.93%). Fifty-four primer pairs were randomly selected for PCR amplification, and 39 of them were successfully amplified. In total, 32 primer pairs were polymorphic in 23 C. camphora germplasms. In addition, unweighted pair-group method with arithmetic means (UPGMA) cluster analysis showed that the similarity coefficients of 23 germplasms ranged from 0.69 to 0.88. At the similarity coefficient of 0.70, these germplasms could be classified into 3 major groups. Finally, 6 primer pairs (c117334, c90845, c108367, c115142, c49114 and c101928) were selected to construct the fingerprints, which could effectively distinguish the 3 new varieties from their offsprings and other germplasms from different geographical sources. The results could provide technical support and theoretical basis for new variety identification, intellectual property protection and molecular assisted breeding of C. camphora.
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Received: 28 November 2022
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Corresponding Authors:
*huanghh@zafu.edu.cn
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