1.Identification and expression analysis of apple PDHB-1 gene family.
Jinghua YANG ; Ju GAO ; Wenfang LI ; Ji LIU ; Jiaxing HUO ; Zhenshuo REN ; Long LI ; Baihong CHEN ; Juan MAO ; Zonghuan MA
Chinese Journal of Biotechnology 2023;39(12):4965-4981
Pyruvate dehydrogenase E1 component subunit beta-1 (PDHB-1) is a gene encoding the β-subunit of pyruvate dehydrogenase complex, which plays an important role in fruit acid accumulation. The aim of this study was to investigate the evolution characteristics of apple PDHB-1 family and its expression in apples with different acid contents. Bioinformatics analysis was performed using databases including NCBI, Pfam and software including ClustalX, MEGA, and TBtools. By combining titratable acid content determination and quantitative real-time PCR (qRT-PCR), the expression of this family genes in the peel and pulp of apple 'Asda' and 'Chengji No.1' with different acid content were obtained, respectively. The family members were mainly located in chloroplast, cytoplasm and mitochondria. α-helix and random coil were the main factors for the formation of secondary structure in this family. Tissue-specific expression profiles showed that the expression of most members were higher in fruit than in other tissues. qRT-PCR results showed that the expression profile of most members was consistent with the profile of titratable acid contents. In the peel, the expression levels of 14 members in 'Asda' apples with high acid content were significantly higher than that in 'Chengji No.1' apples with low acid content, where the expression difference of MdPDHB1-15 was the most significant. In the pulp, the expression levels of 17 members in 'Asda' apples were significantly higher than that in 'Chengji No.1' apples, where MdPDHB1-01 was the most highly expressed. It was predicted that PDHB-1 gene family in apple plays an important role in the regulation of fruit acidity.
Malus/metabolism*
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Fruit/genetics*
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Protein Structure, Secondary
2.Characteristics and phylogenetic analysis of chloroplast genome of a new type of fruit Rubus rosaefolius.
Yongfei WU ; Xuelian YANG ; Xia WANG ; Li YAN ; Wanping ZHANG
Chinese Journal of Biotechnology 2023;39(7):2939-2953
The genomic DNA of Rubus rosaefolius was extracted and sequenced by Illumina NovaSeq platform to obtain the complete chloroplast genome sequence, and the sequence characteristics and phylogenetic analysis of chloroplast genes were carried out. The results showed that the complete chloroplast genome of the R. rosaefolius was 155 650 bp in length and had a typical tetrad structure, including two reverse repeats (25 748 bp each), a large copy region (85 443 bp) and a small copy region (18 711 bp). A total of 131 genes were identified in the whole genome of R. rosaefolius chloroplast, including 86 protein coding genes, 37 tRNA genes and 8 rRNA genes. The GC content of the whole genome was 36.9%. The genome of R. rosaefolius chloroplast contains 47 scattered repeats and 72 simple sequence repeating (SSR) loci. The codon preference is leucine codon, and the codon at the end of A/U is preferred. Phylogenetic analysis showed that R. rosaefolius had the closest relationship with R. taiwanicola, followed by R. rubraangustifolius and R. glandulosopunctatus. The chloroplast genome characteristics and phylogenetic analysis of R. rosaefolius provide a theoretical basis for its genetic diversity research and chloroplast development and utilization.
Phylogeny
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Rubus/genetics*
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Genome, Chloroplast
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Fruit/genetics*
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Codon/genetics*
3.Identification of laccase gene family members in peach and its relationship with chilling induced browning.
Kang WANG ; Minjie YANG ; Siyi WU ; Qingli LIU ; Shifeng CAO ; Wei CHEN ; Liyu SHI
Chinese Journal of Biotechnology 2022;38(1):264-274
The laccase (PpLAC) gene family members in peach fruit were identified and the relationship between their expression pattern and chilling induced browning were investigated. The study was performed using two varieties of peaches with different chilling tolerance, treated with or without exogenous γ-aminobutyric acid (GABA) during cold storage. Twenty-six genes were screened from the peach fruit genome. These genes were distributed on 6 chromosomes and each contained 5-7 exons. The PpLAC gene family members shared relatively similar gene structure and conserved motifs, and they were classified into 7 subgroups based on the cluster analysis. Transcriptome sequencing revealed that the expression levels of PpLAC7 and PpLAC9 exhibited an increasing pattern under low temperature storage, and displayed a similar trend with the browning index of peach fruit. Notably, GABA treatment reduced the degree of browning and inhibited the expression of PpLAC7 and PpLAC9. These results suggested that PpLAC7 and PpLAC9 might be involved in the browning of peach fruit during cold storage.
Food Storage
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Fruit/genetics*
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Laccase/genetics*
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Prunus persica/genetics*
4.Genetic diversity and genetic structures of original plants of Viticis Fructus:an SSR markers-based analysis.
Yi-Qi SUN ; Lu-Ying ZHAO ; Bo ZHU ; Min JIA ; Qiao-Yan ZHANG ; Lu-Ping QIN
China Journal of Chinese Materia Medica 2021;46(15):3824-3831
The present study aimed to provide the protection strategies for wild germplasm resources of original plants of Viticis Fructus and a theoretical basis for the sustainable use of Viticis Fructus. The genetic diversity and genetic structures of the 232 indivi-duals in 19 populations of Vitex rotundifolia and V. trifolia were analyzed by eight SSR markers with tools such as Popgene32, GenAlex 6.502, and STRUCTURE. Bottleneck effect was detected for the population with more than 10 individuals. The results indicated that 42 and 26 alleles were detected from the populations of V. rotundifolia and V. trifolia, respectively, with average expected heterozygo-sities of 0.448 6 and 0.583 9, which are indicative of low genetic diversity. AMOVA revealed the obvious genetic variation of V. rotundifolia and V. trifolia within population(84.43%, P<0.01; 60.37%, P<0.01). Furthermore, in eight SSR loci, six from V. rotundifolia populations and two from V. trifolia populations failed to meet Hardy-Weinberg equilibrium expectations(P<0.05), which confirmed that the populations experienced bottleneck effect. As assessed by Mantel test, geographical distance posed slight impacts on the genetic variation between the populations of V. rotundifolia and V. trifolia. Principal component analysis(PCA) and STRUCTURE analysis demonstrated evident introgression of genes among various populations. The original plants of Viticis Fructus were confirmed low in genetic diversity and genetic differentiation level. Therefore, the protection of wild resources of original plants of Viticis Fructus should be strengthened to ensure its sustainable use.
Alleles
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Fruit/genetics*
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Genetic Variation
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Geography
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Microsatellite Repeats
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Vitex/genetics*
5.Evaluation of genetic diversity of ginseng fruit color germplasm resources: based on SSR analysis.
Ping QIAO ; Yi-Heng WANG ; Jia-Hui SUN ; Chuan-Zhi KANG ; Lan-Ping GUO ; Lu-Qi HUANG
China Journal of Chinese Materia Medica 2022;47(8):2158-2164
Illumina Xten was employed for shallow sequencing of Panax ginseng(ginseng) samples, MISA for screening of SSR loci, and Primer 3 for primer design. Polymorphic primers were screened from 180 primers. From the successfully amplified polymorphic primers, 15 primers which featured clear peak shape, good polymorphism, and ease of statistics were selected and used to evaluate the genetic diversity and germplasm resources of 36 ginseng accessions with different fruit colors from Jilin province. The results showed that red-fruit ginseng population had high genetic diversity with the average number of alleles(N_a) of 1.031 and haploid genetic diversity(h) of 0.172. The neighbor-joining cluster analysis demonstrated that the germplasms of red-fruit and yellow-fruit ginseng populations were obviously intermixed, and pick-fruit ginseng germplasms clustered into a single clade. The results of STRUCTURE analysis showed high proportion of single genotype in pick-fruit ginseng germplasm and abundant genotypes in red-fruit and yellow-fruit ginseng germplasms with obvious germplasm mixing. AMOVA revealed that genetic variation occurred mainly within populations(62.00%, P<0.001), and rarely among populations(39%, P<0.001), but homogenization was obvious among different populations. In summary, pink-fruit ginseng population may contain rare genotypes, which is the basis for breeding of high-quality high-yield, and multi-resistance varieties, genetic improvement of varieties, and sustainable development and utilization of ginseng germplasm resources.
Fruit/genetics*
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Genetic Variation
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Microsatellite Repeats
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Panax/genetics*
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Plant Breeding
6.Isolation and Characterization of Dikaryotic Mutants from Pleurotus ostreatus by UV Irradiation.
Joong Ho JOH ; Beom Gi KIM ; Won Sik KONG ; Young Bok YOO ; Kyo Sun CHU ; Nam Kuk KIM ; Hye Ran PARK ; Bong Gum CHO ; Chang Soo LEE
Mycobiology 2004;32(2):88-94
Protoplasts of the wild type strain of Pleurotus osteatus were mutagenized with UV light, and 3,000 colonies were examined for abnormal mycelial and fruiting phenotypes. Forty one strains displayed variant phenotypes in mycelia and fruiting processes. The variant phenotypes were classified into 6 groups: (1) auxotrophic strains, which are incapable of growing on minimal media and can only grow when provided with their specific requirements; (2) abnormal vegetative strains, which grow very slowly on minimal and complete media; (3) primordiumless strains, which fail to develop to the formation of primordia; (4) maturationless strains, which form primordia, but do not form mature fruiting bodies; (5) specifically colored strains, which have Specific bluish grey or bluish white pileus; (6) poorly spored strains, which fail to produce basidiospore or which produce few spores. These variant strains may be useful in genetic breeding programs and for the studies of fungal development and genetics.
Breeding
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Fruit
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Genetics
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Phenotype
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Pleurotus*
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Protoplasts
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Spores
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Ultraviolet Rays
7.Species identification of Ligustrum lucidum.
Yu-Shuang WANG ; Yuan-Xi JIN ; Kang-Jia LIU ; Chang GUO ; Yi-Heng WANG ; Chao XU ; Zhi-Xiang ZHANG ; Wen-Pan DONG
China Journal of Chinese Materia Medica 2023;48(11):2940-2948
Ligustrum lucidum is a woody perennial plant of genus Ligustrum in family Oleaceae. Its dried fruit has high medicinal value. In this study, the authors evaluated the variability and species identification efficiency of three specific DAN barcodes(rbcL-accD, ycf1a, ycf1b) and four general DAN barcodes(matK, rbcL, trnH-psbA, ITS2) for a rapid and accurate molecular identification of Ligustrum species. The results revealed that matK, rbcL, trnH-psbA, ITS2 and ycf1a were inefficient for identifying the Ligustrum species, and a large number of insertions and deletions were observed in rbcL-accD sequence, which was thus unsuitable for development as specific barcode. The ycf1b-2 barcode had DNA barcoding gap and high success rate of PCR amplification and DNA sequencing, which was the most suitable DNA barcode for L. lucidum identification and achieved an accurate result. In addition, to optimize the DNA extraction experiment, the authors extracted and analyzed the DNA of the exocarp, mesocarp, endocarp and seed of L. lucidum fruit. It was found that seed was the most effective part for DNA extraction, where DNAs of high concentration and quality were obtained, meeting the needs of species identification. In this study, the experimental method for DNA extraction of L. lucidum was optimized, and the seed was determined as the optimal part for DNA extraction and ycf1b-2 was the specific DNA barcode for L. lucidum identification. This study laid a foundation for the market regulation of L. lucidum.
Ligustrum/genetics*
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Seeds
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Fruit
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Polymerase Chain Reaction
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Research Design
8.Complete chloroplast genome of Ligustrum lucidum and highly variable marker identification for Ligustrum.
Yuan-Xi JIN ; Yu-Shuang WANG ; Yong-Wei GAO ; Li-Wei ZHOU ; Yi-Heng WANG ; Qing-Jun YUAN ; Wen-Pan DONG
China Journal of Chinese Materia Medica 2022;47(7):1847-1856
Ligustri Lucidi Fructus, the sun-dried mature fruit of Ligustrum lucidum, is cool, plain, sweet, and bitter, which can be used as both food and medicine, with the effects of improving vision, blacking hair, and tonifying liver and kidney. It takes effect slowly. However, little is known about the genetic information of the medicinal plant and it is still a challenge to distinguish Ligustrum species. In this study, the complete chloroplast genome of L. lucidum was obtained by genome skimming and then compared with that of five other Ligustrum species, which had been reported. This study aims to evaluate the interspecific variation of chloroplast genome within the genus and develop molecular markers for species identification of the genus. The result showed that the chloroplast genome of L. lucidum was 162 162 bp with a circular quadripartite structure of two single-copy regions separated by a pair of inverted repeats. The Ligustrum chloroplast genomes were conserved with small interspecific difference. Comparative analysis of six Ligustrum chloroplast genomes revealed three variable regions(rbcL-accD, ycf1a, and ycf1b), and ycf1a and ycf1b can be used as the species-specific DNA barcode for Ligustrum. Phylogeny analysis provided the best resolution of Ligustrum and supported that L. lucidum was sister to L. gracile. This study clarified the genetic diversity of L. lucidum from provenance, which can serve as a reference for further analysis of pharmacological differences and breeding of excellent varieties with stable drug effects.
Fruit
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Genome, Chloroplast
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Ligustrum/genetics*
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Phylogeny
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Plant Breeding
9.Comparative transcriptome analysis of candidate genes involved in chlorogenic acid biosynthesis during fruit development in three pear varieties of Xinjiang Uygur Autonomous Region.
Hao WEN ; Xi JIANG ; Wenqiang WANG ; Minyu WU ; Hongjin BAI ; Cuiyun WU ; Lirong SHEN
Journal of Zhejiang University. Science. B 2022;23(4):345-351
Pear is one of the main fruits with thousands of years of cultivation history in China. There are more than 2000 varieties of pear cultivars around the world, including more than 1200 varieties or cultivars in China (Legrand et al., 2016). Xinjiang Uygur Autonomous Region is an important pear production region in China with 30 of varieties or cultivars. Pyrus sinkiangensis is the most popular variety, which is mainly distributed in Xinjiang (Zhou et al., 2018). Chlorogenic acid (CGA), p-coumaric acid, and arbutin are the main polyphenols in pear fruit, and their levels show great differences among different varieties (Li et al., 2014). CGA is a potential chemo-preventive agent, which possesses many important bioactivities including antioxidant, diabetes attenuating, and anti-obesity (Wang et al., 2021). Therefore, the specific CGA content of a variety is considered the embodiment of the functional nutritional value of pears.
Chlorogenic Acid
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Fruit
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Gene Expression Profiling
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Pyrus/genetics*
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Transcriptome
10.Identification and characterization of DIR gene family in Schisandra chinensis.
Yu-Qing DONG ; Ting-Yan QIANG ; Jiu-Shi LIU ; Bin LI ; Xue-Ping WEI ; Yao-Dong QI ; Hai-Tao LIU ; Ben-Gang ZHANG
China Journal of Chinese Materia Medica 2021;46(20):5270-5277
Dirigent(DIR) proteins are involved in the biosynthesis of lignin, lignans, and gossypol in plants and respond to biotic and abiotic stresses. Based on the full-length transcriptome of Schisandra chinensis, bioinformatics methods were used to preliminarily identify the DIR gene family and analyze the physico-chemical properties, subcellular localization, conserved motifs, phylogeny, and expression patterns of the proteins. The results showed that a total of 34 DIR genes were screened and the encoded proteins were 156-387 aa. The physico-chemical properties of the proteins were different and the secondary structure was mainly random coil. Half of the DIR proteins were located in chloroplast, while the others in extracellular region, endoplasmic reticulum, cytoplasm, etc. Phylogenetic analysis of DIR proteins from S. chinensis and the other 8 species such as Arabidopsis thaliana, Oryza sativa, and Glycine max demonstrated that all DIR proteins were clustered into 5 subfamilies and that DIR proteins from S. chinensis were in 4 subfamilies. DIR-a subfamily has the unique structure of 8 β-sheets, as verified by multiple sequence alignment. Finally, through the analysis of the transcriptome of S. chinensis fruit at different development stages, the expression pattern of DIR was clarified. Combined with the accumulation of lignans in fruits at different stages, DIR might be related to the synthesis of lignans in S. chinensis. This study lays a theoretical basis for exploring the biological functions of DIR genes and elucidating the biosynthesis pathway of lignans in S. chinensis.
Fruit/genetics*
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Lignans/analysis*
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Phylogeny
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Schisandra
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Sequence Alignment