| Swida wilsoniana is a deciduous tree or shrub of genus Swida of the family Cornaceae.Swida wilsoniana have wide adaptability,excellent materials and strong resistance.They distributed or cultivated in 19 provinces,municipalities and autonomous regions of The Yangtze River flows,except Qinghai,Tibet,Anhui and Shanghai.The oil content of Swida wilsoniana fruit is as high as 34%,which is a high-oil woody oil plant.The oil is rich in a variety of lipa,and it is edible.It can be a national first-grade edible oil after refining,with high nutritional value.It can also be processed into biodiesel after the high-temperature cracking and esterification of the catalyst,which has similar performance to 0#diesel.It has been listed in the key promotion objects of 12 woody oil plants by the State Council of the People's Republic of China.Vigorous development of Swida wilsoniana for oil can alleviate China's energy crisis and improve its energy structure,and at the same time can greatly reduce the import of edible oil in China.At present,China's Swida wilsoniana germplasm resources have not been well collected,preserved,developed and utilized.Phenotypic diversity analysis and SSR marker genetic diversity analysis of Swida wilsoniana can provide a theoretical basis for the construction of core germplasm resources of Swida wilsoniana.In this study,the transcriptome of Swida wilsoniana was sequenced and analyzed,and 34 light bark tree resources were used as test materials.22 phenotypic traits,and 100 EST-SSR primers were tested as research objects to understand the Swida wilsoniana phenotype Correlation between traits and their general classification.The main findings are as follows:(1)The Illumina sequencing platform was used to sequence the transcriptome of Swida wilsoniana,a total of 2193418 clean reads were obtained.After assembly,36839 unigenes were obtained.The Swida wilsoniana transcriptome unigenes are annotated in the NR,Pfam,String,Swissprot,and KEGG databases,accounting for 69.92%,40.11%,37.35%,45.87%,and 28.71%of the total,respectively.Through comparison with the NR database,it was found that the unigene of Swida wilsoniana has the most homologous sequences with Vitis vinifera,followed by Coffea canephora,etc.All unigenes was divided into 3 categories of 58 groups by GO function annotation.And obtained functional classification of 25 categories after KOG annotation.According to the KEGG metabolic pathway annotation,the results showed that 13114 unigenes were involved in 33 metabolic pathways with a total of 5 branches.(2)Using software MISA(version 1.0)search SSRs from 36839 unigenes of Swida wilsoniana.The results shows that 8945 unigenes had SSR,and they account for 24.3%of the total unigenes.A total of 12538 SSR sites were detected from the unigenes,and the frequency of occurrence was about 2514 bp.The SSR length range is from 10 to 329 bp,with an average length of 22.69 bp.The unigenes contained 5,4,3,2 and 1 SSR loci was 11,37,199,1329,8540 unigenes respectively.The SSR richness was in order of Dinucleotide,Mononucleotide,Trinucleotide,Tetranucleotide,Hexanucleotide and Pentanucleotide.There are 210 kinds of repeat units in the SSR,from Mononucleotide to Hexanucleotide has 4,12,60,52,27,55 kinds of repeat units.The number of repetitions of SSR repeat units were mainly in 5-44 times.Four kinds of base sequence with the highest repetition units frequency in different types of SSR is AG/CT,A/T,AT/AT and AC/CT.(3)The optimum combination of Swida wilsoniana SSR-PCR reaction system by orthogonal experiment is:55?(Annealing temperature)+34(Cycle index)+50 ng/?L(Template concentration)+0.5 ?L(Primer quantity).(4)100 primers were verified and screened,and 82 effective primers were obtained,with an effective rate of 82%.The tested primers were used to verify and record 34 samples of Swida wilsoniana.The results showed that the amplification sites were from 9-26,the observed allele number was 9-26,and the average allele number was 20.23.The shanon information index of 34 resources ranges from 0.1492-0.0259,with an average of 0.0988.The largest of it was‘Linchang No.2',followed by 'Laoxing No.1' and 'You B',with a minimum of‘No.49'.The 34 materials were classified into 4 categories by clustering analysis at an average ecliden distance of 15.0 and a clustering coefficient of 33.5.There were 18,11,4 and 1 in the four categories,accounting for 52.94%,32.35%,11.76%,2.94%respectively.(4)The analysis of phenotypic traits results showed that the coefficient of variation of quantitative characters was 4.59%-17.63%,and the degree of variation was as follows:style length>fruit oil content>inflorescence width>leaf area>leaf length>inflorescence length>flesh thickness>ovary length>water content>floret longitudinal diameter>100 fruit weight>floret transverse diameter>leaf width>stamen length>fruit transverse diameter>fruit longitudinal diameter>ovary width>anther length.And the degree of variation of quality characters was as follows:fruit color>leaf shape>fruit shape>leaf color.(5)The results of correlation analysis of phenotypic traits showed that the fruit moisture content was positively correlated with 100 fruit weight and fruit diameter,and significantly positive correlated with leaf length.The oil content of fruit was negatively correlated with fruit diameter,leaf length,leaf area,leaf shape and inflorescence width,and significantly positive correlation with the transverse diameter of flowers.The weight of 100 fruit weight was significantly positive correlated with the transverse diameter and stamen length.The pulp thickness was negatively correlated with leaf shape,and it was positively correlated with the longitudinal diameter of flowers.The longitudinal diameter of fruit was significantly positive correlation with the transverse diameter of fruit,positively correlated with leaf length and leaf area,and the width of inflorescence was negatively correlated.Fruit diameter was negatively correlated with fruit shape.Fruit color was significantly negatively correlated with inflorescence width and style length,and positively correlated with stamen length.Fruit shape was negatively correlated with anther length,etc.(6)The Phenotypic cluster analysis of 34 resources results showed that they were clustered into four categories when the average eclidean distance was 5.0 and the clustering coefficient was 18.538.Category I has 19 resources,Category II has 9 resources,Category III has 5 resources,Category IV has only one resource,accounting for 55.88%,26.47%,14.71%and 2.94%of the total resources,respectively.The results of SSR molecular marker clustering are as follows:all the resources were clustered into four categories when the average eclidean distance is 15.0 and the clustering coefficient was 33.5.There were 19,10,4 and 1 resources respectively,which account for 55.88%,29.42%,1 1.76%and 2.94%.The above research results can provide a theoretical basis for the construction of core germplasm of Swida wilsoniana germplasm resources,and are applied to the collection,preservation and exploitation of Swida wilsoniana germplasm resources. |
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