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| Development and Application of SSR Loci from Functional Genes Involved in Wood Formation in Cunninghamia lanceolata |
| HANG Yun1, YU Jin-Jian1, ZHOU Shi-Shui2, CHENG Jian-Hong1, HUANG Hua-Hong1, *, LIN Er-Pei1, TONG Zai-Kang1 |
1 State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Lin'an 311300, China;
2 Kaihua Forest Farm of Zhejiang, Kaihua 324300, China |
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Abstract To provide an important tool for molecular marker-assisted breeding of wood quality traits, a new nuclear genome SSR marker was developed from functional genes related to Cunninghamia lanceolata wood formation. The SSR locus was detected through analyzing the sequences of 10 genes involved in the formation of C. lanceolata. The availability and polymorphism of the designed SSR primers were examined by capillary electrophoresis. Linkage disequilibrium of pairwise loci and the association analysis between developed SSR loci and wood density were performed by using TASSEL (trait analysis by association, evolution and linkage) GLM (general linear model) program. A total of 15 SSR loci were identified in 7 functional genes. There were di-, tri-, tetra- and hexa- nucleotide repeats, with trinucleotide SSRs being the most frequent,accounting for 46.7% of the total loci. Fourteen pairs of PCR amplification primers were designed according to conserved sequences on both sides of SSR loci. Among them, 7 pairs of primers showed polymorphism. A total of 644 SSR loci combinations were found to have a certain degree of linkage disequilibrium (D'≠ 0), with D' values of 183 combinations greater than 0.5. The association analysis revealed that the CLSSR4 locus from the 5' untranslated region (5' UTR) of ClCesA3 (C. lanceolata cellulose synthase 3) was extremely significantly associated with wood density (P<0.01). The wood density variance explained by this locus was 26.07%, and the corresponding alleles of A201 and A205 showed the positive and negative phenotypic effects, respectively. In this study, seven polymorphic SSR primer pairs were developed from genomic sequence of functional genes, and a SSR locus derived from the 5' UTR of ClCesA3 gene was significantly associated with wood density of C. lanceolata. These results will provide an important tool for molecular marker-assisted improvement of C. lanceolata wood quality.
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Received: 25 June 2018
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Corresponding Authors:
* , huanghh@zafu.edu.cn
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