• Volume 20,Issue 1,2019 Table of Contents
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    • >Review
    • Theory and Practice of the Safe Conservation of Crop Germplasm Resources in China

      2019, 20(1):1-10. DOI: 10.13430/j.cnki.jpgr.20180905001

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      Abstract:China has abundant crop germplasm resources. At present, 340 kinds of crops have been collected and a total of 500,000 accessions have been conserved. How to ensure the safe conservation of germplasm resources is the responsibility and mission of the National Crop Genebank of China. Since the Ninth Five-Year Plan, the National Crop Genebank has systematically studied the theories and technologies of the safe conservation of germplasm. We have made breakthroughs on the key technologies of monitoring and early warning, regeneration and in vitro preservation. Based on the above research, we have created a technical system for the safe conservation of crop germplasm resources in China, which has been applied in the nationwide. The technical system would maximum prolong the safe longevity of germplasm, monitor and warn which need to be regenerated in Genebank, for avoiding the loss of germplasm resources due to the loss of vigor and genetic integrity. Therefore, it may provide a reliable guarantee for the safe conservation and effective utilization of crop germplasm resources in China.

    • Innovation and Utilization of Characteristic Germplasm for Bast Fiber Crops

      2019, 20(1):11-19. DOI: 10.13430/j.cnki.jpgr.20180913001

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      Abstract:This report summarizes the achievements of collection, conservation, identification, innovation, distribution and utilization of bast fiber crop germplasms in China in the past five years. The mapping of important agronomic genes and deciphering of their molecular mechanism were described. The current status and future prospective of characteristic medicinal hemp and jute with functional leaf and high adsorption capacity for heavy metals are also reviewed. We believe that the theoretical and applied research of bast fiber germplasm have potential for serving the industries, the regional economic development, the targeted poverty alleviation and the rural revitalization.

    • >Research Articles
    • Test for Ear Rot Resistance Against Fusarium verticillioides and Fusarium graminearum in Imported Maize Germplasm

      2019, 20(1):20-25. DOI: 10.13430/j.cnki.jpgr.20180530001

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      Abstract:A total of 177 maize germplasm accessions, which were imported from the countries of USA, Canada, Romania, Germany, France and Netherlands, were evaluated for resistance against ear rot causing agent of Fusarium verticillioides (FV) and Fusarium graminearum (FG) with double toothpick inoculation technique in two calendar years (2016-2017). Out of that, two accessions (1.13% of total tested accessions) were highly resistant (HR) to FV, and 38 accessions (21.47%) were resistant (R). 75, 48 and 14 accessions showed moderate resistance (MR, 42.37%), susceptible (S, 27.12%) and highly susceptible (HS, 7.91%), respectively. Upon inoculation of FG, 12 and 66 accessions were resistant (R, 6.78%) and moderately resistant (MR, 37.29%), respectively. Meanwhile, 64 and 35 accessions were susceptible (S, 36.16%) and highly susceptible (HS, 19.77%) , respectively. No accessions with highly resistant (HR) to FG was not found. Attractively, twelve accessions with resistant to both FV and FG were identified, and they might serve as resistance resource used in maize breeding for ear rot resistance.

    • Genetic Diversity of Old Tea Plant Resources in Jiuan City of Guizhou Province, Using Genome-Wide SNPs

      2019, 20(1):26-36. DOI: 10.13430/j.cnki.jpgr.20180529001

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      Abstract:A large number of old tea plant resource is widely distributed in Jiuan city of Guizhou province, China. The genetic relationship of these tea plant resources still remain unclear. In this study, one-hundred old tea plant samples, which were collected from five different regions of Jiuan city, were subjected for diversity analysis by genotyping by sequencing (GBS). A total of 548,597 high-quality SNPs were identified in this collection, and the variation of these SNPs were annotated. By using these genome-wide SNPs, the phylogenetic tree, the principal component analysis (PCA) and population structure were conducted. A similar pattern on genetic relationship was revealed by three approaches. These samples were assigned into three groups and the samples collected in the same/similar area were clustered in each group. Thus, our results suggested a genetic difference of samples in the Guchayuan (G), relative to the samples from other four regions, which likely shared the same and unique parental origin.

    • Genetic Diversity and Structure Analysis in Psathyrostachys Population Using SSR Markers

      2019, 20(1):48-59. DOI: 10.13430/j.cnki.jpgr.20180612002

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      Abstract:Genetic diversity and population structure analysis of 171 wild and cultivated Psathyrostachys individual germplasms from different countries and regions were conducted by 38 pairs of SSR primers. The result by using the effective primers revealed a total of 308 loci among Psathyrostachys individuals, with 275 polymorphic loci and a polymorphism percentage of 89.29%. The genetic similarity coefficient of the tested materials ranged from 0.537 to 0.899 with an average of 0.742. These accessions were divided into three groups by population structure analysis, principal component analysis and NJ clustering. The genetic similarity coefficient analysis showed that the distribution of Psathyrostachys individuals was more concentrated. The classification using three methods generally revealed three different groups of Psathyrostachys materials, despite that there were slightly different in the divisions of mixed groups. Since these Psathyrostachys individuals in each group showed different on genetic background, no clear correlation between geographic distribution and genetic diversity was observed.

    • Analysis on Genetic Diversity for Part of Japonica Resources in Heilongjiang Province

      2019, 20(1):60-68. DOI: 10.13430/j.cnki.jpgr.20180725002

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      Abstract:The objective of this paper is to clarify the genetic relationship of japonica rice resources from different breeding units of Heilongjiang province. Genetic diversity and population structure of 104 japonica rice accessions, which were collected from eight different breeding units, were analyzed by 45 pairs of simple sequence repeat (SSR) molecular markers in this study. The results showed that all selected markers were applicable in clarification of the genetic diversity, yielding a total of 154 alleles. The alleles (Na) per locus ranged from 2 to 8 with the mean value of 3.42. The gene diversity indices (H) varied from 0.02 to 0.77 with an average of 0.38. The polymorphism information content (PIC) was changed from 0.02 to 0.73 with the average of 0.34. Furthermore, the genetic diversity of rice varieties from Heihe Branch of Heilongjiang Academy of Agricultural Sciences was the most abundant, while the Northeast Agricultural University was narrow, with the polymorphism information content (PIC) of 0.33 and 0.18, respectively. By UPGMA Cluster and genetic structure analysis, the rice cultivars from Heihe Branch of Heilongjiang Academy of Agricultural Sciences represented genetic differentiation and the other units, while a closer relationship between Rice institute of Heilongjiang Agricultural Reclamation Academy and Jiamusi Rice Research Institute of Heilongjiang Academy of Agricultural Sciences was observed. Thus, our results suggested a demand of enlarging the genetic basis in breeding for elite varieties in Helongjiang province, by taking use of the genetic resource from other units in future.

    • Genetic Diversity Analysis of Broomcorn Millet (Panicum miliaceum L.) of Shanxi Province Using Microsatellite Markers

      2019, 20(1):69-78. DOI: 10.13430/j.cnki.jpgr.20180718001

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      Abstract:Abstract: Shanxi province of China acts as one of the main producing areas of common millets and also a center of local varieties and farmer germplasm resources. Exploration of genetic diversity in common millet germplasms will provide a scientific basis for studying the origin and evolution of proso millet, which would strengthen gene transfer between varieties of common millet and facilitate the utilization of excellent resources. In this study, we analyzed the genetic differences among 57 common millet accessions from Shanxi province by 27 pairs of SSR polymorphic primers. The statistical analysis were conducted by software PowerMarker 3.25, PopGen 1.32, MEGA 5.0, Structure 2.3.4 and Ntsys2.11. Within this collection, 71 alleles were detected with an average of 2.6296 alleles at each locus. The number of effective allele account for 76.94% of the observed one. The Shannon index of all primers ranged from 0.4140 to 1.0446, with an average of 0.7686, The range of PIC was 0.0601-0.7158, with an average of 0.5667. The values of genetic diversity indexes such as the observed heterozygosity have no much difference between two ecotopes. Basing on Neighbor-joined genetic distance, 57 materials were divided into 3 groups, the group Ⅰ belongs to the Loess Plateau spring & summer-sowing ecotopes, the group Ⅱ belongs to the Loess Plateau spring & summer-sowing and Northeast spring-sowing, the same as the group Ⅲ. The analysis of genetic structure divided 57 common millet accessions into three groups. In terms of the values of Shannon''s information index and PIC, the green group is higher than of the blue and red. The results of two cluster analysis are consistent and they showed a close correlation between geographical region and the method of clustering. Meanwhile the materials in northern of Shanxi province were distinguished from the middle and the southern of Shanxi province by the means of PCA, The result indicated that the genetic background of two ecotopes has obvious differences due to geographical and climatic factors, which is consistent with the result of genetic structure analysis between populations. Thus, the genetic diversity of common millet resources in Shanxi province was abundant.

    • The Genetic Diversity and Population Structure Analysis of Cultivated Species Malus prunifolia(Willd.)Borkh. of Malus Mill.

      2019, 20(1):79-89. DOI: 10.13430/j.cnki.jpgr.20180522002

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      Abstract:Genetic diversity and population structure analysis were carried out on the newly collected germplasm resources of cultivated species Malus prunifolia of Malus Mill. by use of the fluorescent SSR molecular markers. The identification of the genetic diversity and structure in and among populations can provided references for the collection and preservation of germplasm resources of Malus, and for the selection of rootstock breeding. Fingerprint data were constructed using fluorescent SSR molecular markers, genetic diversity was analyzed using GenAlEx 6.501, Neighbour-Joining tree was constructed using POPULATION 1.2, and population structure was analyzed using STRUCTURE 2.3.4. The results showed that 390 polymorphic alleles were detected by 19 pairs of SSR primers, the average allele number was 20.526, and the average effective allele number was 9.399. The average values of heterozygosity and expected heterozygosity were 0.706 and 0.868, respectively. The Shannon index was 2.446. The genetic diversity of Malus prunifolia in this study was higher than the other species of Malus Mill. before. The cluster analysis based on Nei genetic distance can be divided into three groups for 155 accessions at 0.9167. Three groups are close with each other, and most groups are not related to geographical location. The population structure analysis of all accessions was divided into two groups. Clustering results coincide with the population structure. Q value of 150 accessions was higher than 0.6, and these accessions had a relatively single blood relationship.

    • Identification and Bioinformatics Analysis of Maize SAUR Gene Family

      2019, 20(1):90-99. DOI: 10.13430/j.cnki.jpgr.20180707001

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      Abstract:In order to study the small auxin-up RNA (SAUR) family of maize, this study identified by the genome-wide prediction approach 91 SAUR genes which named ZmSAURs, and analyzed the gene structure, amino acid characteristics, chromosomal location and genetic evolutionof ZmSAURs. The results showed that, the SAUR gene family were unevenly distributed on the chromosome, while the chromosome 2 up to 22 ZmSAURs. The amplification pattern of the gene was dispersed replication and fragment replication. The SAUR gene family has a relatively conserved structure, e.g. a conserved RNA DNA structure and the 3D structure of the deduced proteins that generally contained three alpha helices and three beta sheets. The phylogenetic tree analysis revealed 9 branches of SAUR proteins among species. As expected, SAURs in corn and relative millet (Setaria italic) were clustered together. Thus, this information provided by this work might be useful in future functional characterization of the maize SAUR gene family.

    • Evaluation on Drought Resistance of Winter Wheat Cultivars in Xinjiang

      2019, 20(1):100-112. DOI: 10.13430/j.cnki.jpgr.20180507001

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      Abstract:The comprehensive evaluation of drought resistance at the germination stage was conducted in winter wheat cultivars from Xinjiang, which provided the theoretical basis for the evaluation and selection of drought resistant wheat varieties during the germination period. In this study, two methods that possessed 20% PEG-6000 treatment and marker-assisted selection (MAS) with the functional markers TaNRX-a1 and TaNRX-b1 located on the 5BS chromosome, were used for adjusting drought resistance in 83 Xinjiang winter wheat cultivars at the germination stage. Seven indexes including germination rate, germinating energy, germination index, root number, root length, seedling height and coleoptile length were measured, and the results suggested that the seven indexes were decreased by 30.86%, 23.91%, 31.37%, 0.43%, 31.07%, 50.27% and 6.60%, respectively. The allele frequency of TaNRX-a and TaNRX-b in 83 winter wheat cultivars accounted for 83.1% and 16.9%, respectively. The TaNRX-a allele with high drought resistance during seed germination is predominant, while this allele was observed 93.8%, 82.1% and 79.5% of local varieties, introduced varieties and improved varieties, respectively. By applying the weighted membership function method together with MAS for TaNRX-a, the cultivars Chang dong 5, Hanyangmai, Kayimuhong, Nawatiran, and Gaojiasuo represented higher drought resistance. The correlation analysis showed, that both the root length and the coleoptile length were highly correlated with D value (0.557 and 0.516), and they could be served as a sign in selecting for drought resistance of wheat. Thus, taking use of phenotype datasets and molecular markers, this work suggested a way to effectively evaluate the drought resistance of wheat during germination stage.

    • Correlation Analysis of Yield per Plant and Agronomic Traits of Breeding Parents in Brassica napus L.

      2019, 20(1):113-121. DOI: 10.13430/j.cnki.jpgr.20180705001

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      Abstract:Rapeseed acts as an important oil crop in China and breeding for high yield is of great interest. Analyzing agronomic traits of rapeseed germplasm can provide foundational information for selecting excellent parental materials, which will be subjected for making crosses, e.g. to improve important traits in rapeseed. In the present study, 213 breeding lines of B. napus L., which were collected from the main rapeseed production area of the Yangtze River, was used for exploring the relationship between 10 agronomic traits and yield per plant, under the climatic conditions in Chengdu Flat. The statistical analysis including simple correlation analysis, path analysis, multiple regression analysis and principal component analysis, were conducted. The results showed the great divergence on the agronomic traits of rapeseed resources from different regions. The yield per plant was positively correlated to plant height, the effective length of main inflorescence, the number of first effective branch, the number of effective silique on main inflorescence, the length of silique, seeds per silique, and 1000-seed weight, while negatively correlated to the period of vegetative growth. The yield per plant was directly contributed by three factors including plant height, seeds per silique and the number of first effective branches. The indirect effect on plant yield was the effective length of main inflorescence > the number of effective silique on main inflorescence > branch height. By stepwise regression analysis, the yield per plant was mainly determined by plant height, seeds per silique, branch height, and the number of first effective branches. Based on the phenotypic variation using the principal component analysis, these genotypes were grouped into four major categories, consisting of plant height related factors, growth period related factors, yield related factors and plant type related factors. Taken together, the yield per plant in the ecological region of Chengdu Flat was largely contributed by plant height, grain number per pod, primary branch height, and number of primary effective branch. These results would provide a valuable information for selecting elite parental lines in rapeseed breeding.

    • Genetic Diversity and Fingerprint Analysis of Mungbean Varieties from China Based on InDel Markers

      2019, 20(1):122-128. DOI: 10.13430/j.cnki.jpgr.20180524002

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      Abstract:Mungbean is a kind of crop with strong resistance, and wide adaptability. The genetic diversity analysis and fingerprint analysis of all samples were detected using 19 polymorphic and stable core markers. The effective alleles of the InDel markers were between 1.100 and 1.996, with an average of 1.750. The expected heterozygosity (He) was between 0.0918~0.5049, with an average of 0.421. The variation of Nei"s gene diversity index was between 0.0907~0.4989, with an average of 0.416. The value of the polymorphic information content (PIC) was between 0.0866~0.3744, and 0.25 < 0.50 was the medium polymorphic. The average PIC content of InDel markers used in this experiment was 0.325. Using UPGMA to construct the clustering tree of 42 varieties of mungbean germplasm resources and the result showed that, 42 mungbean varieties (lines) could be divided into 2 groups while the genetic similarity coefficient is 0.55. Genetic analysis showed that the germplasm resources of the domestic mungbean have been isolated from the effect of regional isolation. By using 19 InDel markers, the DNA fingerprint of 42 cultivars of mungbean was constructed and 42 varieties of mungbean could be distinguished and accurately identified. The bands of InDel markers are easy to read in this study and they are suitable for DNA fingerprinting. Our results provide scientific basis for the identification of varieties, the protection of varieties of mungbean and provenance management.

    • Genetic Diversity Analysis of Sinopodophyllum hexandrum Germplasm Accessions Using ISSR Markers

      2019, 20(1):129-136. DOI: 10.13430/j.cnki.jpgr.20180504001

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      Abstract:: Objectiv To analyze the genetic polymorphism and relationship of Sinopodophyllum hexandrum from Gansu and Qinghai province in China, and to offer some information for the domestication and protection of wild Sinopodophyllum hexandrum. Methods ISSR primers were used to analyze the genetic diversity of 11 populations of Sinopodophyllum hexandrum. Cluster analysis of the unweighted pair-group method with arithmetic average (UPGMA) and do Mantel test with geographic distance, out based on the molecular data obtained, and a dendrogram was constructed. Results Totally 155 alleles were amplified using the 11 ISSR primers. The number of alleles in per primer range was 10-17, with an average of 14.1 and 1320 polymorphic locis. The average percentage of polymorphic loci (PPL) was 65.50%. The Nei’s gene diversity (H) and Shannon’s information index (I) were 0.2101and 0.3191. The genetic distances range was 0.0316-0.3769,and Mantel test p=0.4220. Conclusion The genetic polymorphism among 13 populations of Sinopodophyllum hexandrum is relatively abundant and the gene flow within the populations were very rich, the genetic differentiation within the population is significantly greater than the differentiation between populations and there is no relationship between genetic distance.

    • Field Collection and Genetic Diversity Analysis of Pea ( Pisum sativum L. ) Germplasm Resource in Chongqing

      2019, 20(1):137-145. DOI: 10.13430/j.cnki.jpgr.20180528001

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      Abstract:Under the frame of the Third National Survey and Collection Action on Crop Germplasm Resources, the Chongqing Project Group collected 56 pea germplasm accessions from 19 counties of Chongqing from 2015 to 2017. These accessions were found at wide geographic regions with different altitude. We investigated fourteen agronomic traits that were subjected for genetic diversity analysis. The abundant genetic diversity at the agronomic traits was observed, including the qualitative trait pod shape and the quantitative trait plant height with the highest diversity index, as well as the branching number that is with the highest coefficient variation (CV). These accessions were divided into 5 groups by clustering analysis: (1) the accessions with erect or early-mature; (2) the accessions with elite grain edible; (3) the accessions with edible leaf and edible pod; (4) the accessions with high-yield potential and medium-size grain; (5) the accessions with large gain and pod edible. Thus, these pea accessions represented phenotypic variations on pod-eating, leaf-eating and grain characteristics, and this germplasm resource might be valuable in breeding for new pea varieties in Chongqing.

    • Investigation and Analysis of Medicinal Plant Resources in Longling County of Yunnan Province

      2019, 20(1):146-158. DOI: 10.13430/j.cnki.jpgr.20180329001

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      Abstract:Under a frame of the fourth national Traditional Chinese Medicine (TCM) resource survey, we conducted a two-year survey (2015 and 2016) on the geographical distribution of the medicinal plant resources in Longling county of Yunan province. According to the guidelines, wild?and cultivated medicinal?plant?resources were investigated, and traditional medicine knowledge and market was surveyed. In Longling, the field survey was conducted at 37 different locations of 10 different sub-district of Longling county. The medicinal plants and its specimens?were collected with data of interview from the local and assessing by investigation team. The herbariums were numbered to be 653#. Interestingly, we found that 606 species belonging to 437 genus, 142 families, were shown to be traditional medicine. For example. we identified 84 important medicinal materials, comprising of three types: (1) 48 species of origin plants of wild herbs, (2) 36 species of origin plants of cultivation herbs, and (3) 24 species that are listed as national rare and endangered medicinal plants. >30 species of medicinal plants were found in Longling county, including wild medicinal plants clubmosses, cogongrass rhizome, golden buckwheat, puerarin, dichroa, huang jing, rhizoma arisaematis, etc., and cultivated medicinal plants Caulis dendrobii, amomum, polyphylla, Paris polyphylla, gentiana, aconite, Radix dipsaci, notoginseng, spreading hedyotis herb, salvia miltiorrhiza, platycodon grandiflorum, Sculellaria barbata, Rhizoma polygonati, angelica, CARDS, etc. Out of that, Caulis dendrobii, amomum and Paris polyphylla Comparing were found to be cultivated with large area. Furthermore, a large number of valuable medicinal plants were found to be existed and widely distributed Longling county. Thus, the findings of this study might provide insights and germplasm resources that potentially support sustainable development of medicinal plant resources of Longling county.

    • Characterization of a Candidate Transcription Factor WRKY55 that Associated with the Infection of Barley yellow mosaic virus

      2019, 20(1):159-167. DOI: 10.13430/j.cnki.jpgr.20180529002

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      Abstract:Barley yellow mosaic virus disease, caused by Barley yellow mosaic virus (BaYMV) and Barley mild mosaic virus (BaMMV), is a major viral disease in barley planting regions of Europe and East Asia countries. In the field, this disease is transmitted by the soil-borne fungus Polymyxa graminis via inoculating the roots. Here, by unlocking the transcriptome datasets, we identified a differentially expressed gene HORVU1Hr1G069640 (referred WRKY55) that was significantly up-regulated after infection with BaMMV. Consistently, by analyzing its expression in a group of naturally-inoculated genotypes, a significantly higher expression of WRKY55 was observed in susceptible relative to resistant genotypes. This gene contains an 870 bp of the coding sequence with a conserved WRKY domain between nucleotides 415’-594’. Phylogenetic analysis revealed an independent branch of this gene, differed to the sequence homologous genes. This gene was highly expressed in roots and young tissues, but rarely expressed in other tissues. By Agrobacterium-mediated transformation into Nicotiana benthamiana, the WRKY55-GFP fusion protein was found in the whole cell. In yeast, no trans-activation activity of WRKY55 was detected and no physical interaction between WRKY55 and either susceptibility factors eIF4E or PDIL5-1 were observed. This work identified a BaMMV-infected up-regulated gene WRKY55, which possibly was involved in infection of BaMMV/BaYMV.

    • Genome-wide Characterization of the LBD Gene Family in Radish

      2019, 20(1):168-178. DOI: 10.13430/j.cnki.jpgr.20180503002

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      Abstract:LBD gene family is a kind of plant-specific transcription factors and plays very important role in plant development. In this study, 59 LBD genes distributed on 9 chromosomes of radish genome were identified. The radish LBD genes have simple structures with the intron numbers being less than 3. These LBD genes could be classified into 2 classes (Class I and Class II) according to the phylogeny relationship, including 50 and 9 members, respectively. The number of radish LBDs distributed on different chromosomes has significantly difference, 18 members distributed on chromosome 1 while only one member distributed on chromosome 7 and 8, respectively. The expression patterns of radish LBD genes in different tissues and development stages showed spatial and temporal differences, uncovered their potential diverse function in radish development. These results will be helpful for the functional analysis of the radish LBD genes family.

    • Development and Functional Analysis of Peanut Insertion and Deletion (InDel) Markers Based on Genotyping-by-sequencing (GBS)

      2019, 20(1):179-187. DOI: 10.13430/j.cnki.jpgr.20180517003

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      Abstract:Insertion/deletion (InDel) is the second largest molecular markers, which provide the valuable reference for population genetics and marker-assisted selection study. Peanut is one of the most important oil crops worldwide. The whole genome sequence of cultivated peanut has been released and therefore provided the important genome reference for functional genomic studies. In this study, by taking use of the genotyping-by-sequencing datasets on 169 peanut core collections, we identified 10,401 InDels with the size difference of 1 to 14 bp. The highest number of InDels was observed on chromosome Arahy.16, while the lowest number was found on Arahy.08. Although only 1,167 Indels were located in coding regions, obviously these InDels might show the specific functions, including catalytic activity and binding compared with other regions. The KEGG analysis suggested that InDels were mainly detected in the metabolism process. These InDels explored in this study might serve as the important genetic resources being useful for the further validation of functional genes.

    • Isolation of CpActin Gene from Cucurbita pepo and It's Preliminary Application as an Internal Standard

      2019, 20(1):188-198. DOI: 10.13430/j.cnki.jpgr.20180607001

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      Abstract:Quantitative real-time PCR (i.e., qRT-PCR), as one of nucleic acid quantification technologies used for analyzing gene expression in molecular biology, has provided the advantage of high efficiency, high specificity, good repeatability, high sensitivity and high degree of automation. Therefore, selecting suitable internal reference genes based on the corresponding experimental materials is particularly important for ensuring the accuracy of qRT-PCR analysis. Since the Actin gene expresses consistently without tissue specificity, it has been often used as internal reference for normalizing the gene expression. Previously, we generated the early low-temperature and low-light transcriptome datasets of Cucurbita pepo using transcriptome sequencing (i.e., RNA-seq) technology. Here we identified a 2,543 bp cDNA from this database, with a size of 2,064 bp open reading frame (i.e., ORF) encoding 682 amino acids. This deduced protein showed a theoretical molecular weight of 79.67 kD and a protein isoelectric point (i.e., PI) of 6.28. Wolf Psort analysis indicated that CpActin protein was located in the nucleus, and Motif Scan analysis showed that CpActin protein had the Mid-end and C-end domains of conserved Actin in the position of 207-406 and 558-682 sites, respectively. Cpactin shared 99% identity on amino acids with the homologous proteins from the relatives Cucurbita moschata and Cucurbita maxima, proving that this protein was highly conservative. By using a pair of primers that was designed for amplifying the full-length ORF, we re-generated and verified the CpActin fragment (Genbank ID: MH211008). What’s more, a pair of qRT-PCR primers CpActin-Fq and CpActin-Rq were designed. The analysis showed that the pair primers has high specificity and amplification efficiency, which can be stably expressed in different tissues (i.e., Root, Stem, Flower, Fruit) and different stress treatments [i.e., Normal (25℃, 300 umol&#183;m-2&#183;s-1), Low temperature treatment (4℃, 300 umol&#183;m-2&#183;s-1), Low light treatment (25℃, 80 umol&#183;m-2&#183;s-1), Low temperature and low light treatment (4℃, 80 umol&#183;m-2&#183;s-1), High light treatment (25℃, 2000 umol&#183;m-2&#183;s-1) and High temperature treatment (38℃, 300 umol&#183;m-2&#183;s-1) ] leaves of Cucurbita pepo. And it was suitable for use as a candidate internal reference gene in the study of qRT-PCR expression analysis of Cucurbita pepo. Thus, this study provided a reference gene CpActin being suitable for qRT-PCR analysis of important functional genes of Cucurbita pepo.

    • Study on the Genetic Diversity and Genetic Structure of Cardiocrinum giganteum Based on rpl16 Sequence Analysis

      2019, 20(1):199-206. DOI: 10.13430/j.cnki.jpgr.20180427001

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      Abstract:Cardiocrinum giganteum belongs to the genus Cardiocrinum (Endl.) Lindl. (Liliaceae), which is an important wild plant resource. In this study, the genetic diversity and genetic structure of 10 C. giganteum populations were analyzed by using the rpl16 sequence of the chloroplast DNA in order to provide theoretical basis for its resource protection and utilization. By alignment of 699 bp sequence, 14 mutation sites and 12 haplotypes were identified, with the haplotype polymorphism (Hd) and the total genetic diversity (HT) of 0.604 and 0.660, respectively. The genetic variation among the population (71.5 3%) was higher than the intra-population genetic variation (28.47 %), indicating the population variation largely contributed by different populations. The genetic differentiation among the population was high (Gst = 0.677, Nst = 0.614, Fst = 0.71531) and the gene flow was low (Nm = 0.0995). Mismatch analysis revealed that the population had experienced rapid expansion during the evolution process, which was supported by the neutrality tests (Tajima's D = -1.74368 (P > 0.05); Fu and Li's D = -2.69366 (P < 0.05); Fu and Li's F = -2.79896 (P < 0.05)). The study revealed the abundant genetic diversity and differentiation of C. giganteum particularly among populations, which was mainly related to the characteristics of life history. Thus, this work suggested an in-situ conservation strategy being employed in C. giganteum.

    • The Construction of CsTHCA RNA Interference Vector and Its Genetic Transformation in Cannabis sativa L.

      2019, 20(1):207-214. DOI: 10.13430/j.cnki.jpgr.20180524001

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      Abstract:CsTHCA plays important role in regulating THC contents. In this study, we isolated the complete coding sequence of CsTHCA from Cannabis Sativa L., with an open reading frame of 1638 base pair. The deduced CsTHCA protein contained 545 amino acids. The CsTHCA RNA interference construct was generated through pSKint intermediate vector, which was transformed into the shoot tips of hemp. By analyzing the transcriptional level of CsTHCA by semi-quantitative RT-PCR and the THC content by thin-layer chromatography and SPE-HPLC, the reduction of CsTHCA expression and THC content was observed in transgenic lines relative to wild type. Thus, our results supported a positive role of CsTHCA in modulating THC content in hemp.

    • Phenotypic Identification and Genetic Analysis of A Novel Tomato Yellow Green Leaf Mutant

      2019, 20(1):215-220. DOI: 10.13430/j.cnki.jpgr.20180518001

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      Abstract:Plant yellow-green leaf mutant plays an important role in a series of basic studies such as photosynthesis, synthetic metabolic pathway of chlorophyll, genetic differentiation and development of chloroplast, as well as a marker trait to breeding research. In this study, we analyzed the phenotypic characteristics and photosynthesis of a tomato yellow-green leaf mutant that was generated by chemical mutagenesis. We found that the yellow-green phenotype of mutant (mu) became visible in the first true leaf. The mu plant showed dwarfism, the reduction of chlorophyll content and net photosynthetic rate in relative to that of the wild type M82. By analyzing the ultra-structure of chloroplast with transmission microscope, the chloroplast thylakoid was impaired development in the mu mutant. By genetic analysis between mu and M82, the segregation ratio (3:1) was observed in the seedlings with normal and yellow leaves in a F2 population, supporting a single recessive gene underlying the mutated yellow-green phenotype. Thus, this study identified a chloroplast-abnormal mutant that might be useful in exploring the signaling in nucleus and chloroplast, for example, by isolating the target gene controlling the yellow-green phenotype of leaves.

    • Genetic Diversity Analysis of Foxtail Millet Varieties (Setaria italica L.) Released from 1980s to 2010s in Summer Sowing Region of North China Using SSR Markers

      2019, 20(1):221-228. DOI: 10.13430/j.cnki.jpgr.20180525001

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      Abstract:The genetic diversity of 20 representative foxtail millet varieties, which were released in summer sowing region of North China since 1980s, were analyzed using 49 pairs of SSR markers. These SSR markers unlocked 142 alleles with a mean of 2.96 alleles per locus. The polymorphism information content (PIC) ranged from 0.0904 to 0.6896, with an average of 0.4168. The mean genetic distance was 0.4045 with a range from 0.0173 to 0.9000. These foxtail millet varieties were classified into 3 groups based on SSR assay. The variety Gufeng No.1 formed an independent group, suggesting a higher genetic distance of this variety to others. By investigating the genetic distances of foxtail millet cultivars that were released in different decades, the higher genetic diversity was observed in older varieties (1980s>1990s>2000s>2010s), which indicated a constitutive drop of the genetic variation in foxtail millet cultivars in breeding over last few decades.

    • Analysis of Main Phenotypic Characteristics in Coix L. Germplasm Resources

      2019, 20(1):229-238. DOI: 10.13430/j.cnki.jpgr.20180604003

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      Abstract:The objective of this study attempts to evaluate the diversity of Coix germplasm resources by phenotypic chracters and thus provide basis for parent selection and variety improvement.A total of 22 phenotypic traits were investigated on 108 Coix L. germplasm accessions, which contained two species and four varieties of Coix L. The genetic diversity of 9 descriptive traits (colour of coleoptile, seaf sheath, seeding leaf, stigma, young fruit, presence of involucre, involucre shape and texture) ranged from 0.55~1.65.The variation coefficients of 12 numerical characters (plant height, grain layer, stem diameter, length of leaf, width of leaf, total tillers per plant, productive tillers per plant, nodes on main stem, branching node, 100-fruit weight, involucre length and involucre width) ranged from 13.0%~60.1%. Moreover, most of numerical traits related significantly to each other, and five principle componence were extracted and simplified from the phenotypic traits with a cumulative percentage of 77.4% by principle componence analysis. By cluster analysis, these acceccsions were clarified into three groups, including: group-I, in which these accessions showed dwarf plants, smaller leaf, fewer nodes and lower branching nodes, and in which they were distributed in the higher latitude areas; group-II, in which these accessions were distributed in the lower latitude areas of the middle and lower reaches of Yangtze river, south and southwesr of China and southeast Asia, showing medium plant height, fewer tillers, stem diameters, nodes and branching nodes, leaf sizes there; group-III, in which the accessions were distributed in southwest of China, with the higher plants and thicker stem diameter, larger leaf size, more nodes and higher branching nodes, more tillers and higher biomass. Thus, by taking use of this collection with regional peculiarity and distinct phenotypic variation, this work might provide insight for selecting parental materials being valuable in breeding for elite adlay.

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