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    园艺作物种质资源与遗传育种Horticulture — Genetics · Breeding · Germplasm resources

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    Isolation and Functional Analysis of the bZIP Transcription Factor Gene TaABP1 from a ChineseWheat Landrace
    CAO Xin-you, CHEN Ming*, XU Zhao-shi, CHEN Yao-feng, LI Lian-cheng, YU Yue-hua, LIU Yangna, MA You-zhi
    2012, 12 (10): 1580-1591.   DOI: 10.1016/S1671-2927(00)8691
    Abstract1642)      PDF in ScienceDirect      
    In plants, basic leucine zipper (bZIP) transcription factors play important roles in regulatory processes, including stress response, pathogenic defense and light response as well as organ and tissue differentiation. Chinese wheat landrace Pingyaoxiaobaimai (PYXBM), an original parent of drought tolerant wheat varieties grown in northern China, is significantly tolerant to abiotic stresses such as drought, cold and nutrient deficiencies. In order to isolate key stress-responsive genes and then improve stress tolerances of conventional varieties, a bZIP transcription factor gene was isolated from a cDNA library of drought-treated PYXBM using the in situ plaque hybridization method, and was designated as Triticum aestivum L. abscisic acid (ABA)-responsive element binding protein 1 (TaABP1). It encodes 372 amino acids, and contains three conserved domains (C1-C3) in the N terminal and a bZIP domain in the C terminal which is a typical protein structure for the group member of bZIP family. Transcriptional activation analysis showed that TaABP1 activated the expression of downstream reporter genes in yeast without ABA application. TaABP1 protein fused with green fluorescent protein (GFP) demonstrated that the localization of TaABP1 protein is in the nucleus. Expression pattern assays indicated that TaABP1 was strongly induced by ABA, high salt, low temperature and drought, and its expression was stronger in stems and leaves than in the roots of wheat. Furthermore, overexpression of TaABP1 in tobacco showed significant improvement of drought tolerance. Data suggested that TaABP1 may be a good candidate gene for improving stress tolerance of wheat by genetic transformation and elucidation of the role of this gene will be useful for understanding the mechanism underlying drought tolerance of Chinese wheat landrace PYXBM.
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    QTL analysis of early flowering of female flowers in zucchini (Cucurbita pepo L.)
    QU Shu-ping, YANG Dan, YU Hai-yang, CHEN Fang-yuan, WANG Ke-xin, DING Wen-qi, XU Wen-long, WANG Yun-li
    2023, 22 (11): 3321-3330.   DOI: 10.1016/j.jia.2022.09.009
    Abstract182)      PDF in ScienceDirect      

    Early flowering promotes early maturity, production, and the capacity to counteract biotic and abiotic stresses, making it an important agronomic trait in zucchini.  The present study demonstrated that the zucchini inbred line ‘19’ consistently flowered early, taking significantly fewer days to bloom the first female flower (DFF) than the inbred line ‘113’.  Genetic analysis revealed that DFF, an inheritable quantitative trait, is controlled by multiple genes.  Based on the strategy of quantitative trait locus (QTL) sequencing (QTL-seq) combined with linkage analysis, three QTLs for DFF were identified on chromosomes 4, 11, and 20.  This study used additional F2 populations grown under different environmental conditions for QTL mapping analysis of DFF with insertion/deletion (InDel) markers to validate these results.  Using the composite interval mapping (CIM) method of R/qtl software, we only identified one major locus under all environmental conditions, located in a 117-kb candidate region on chromosome 20.  Based on gene annotation, gene sequence alignment, and qRT-PCR analysis, we found that the Cp4.1LG20g08050 gene encoding a RING finger protein may be a candidate gene for the opposite regulation of early flowering in zucchini.  In summary, these results lay a foundation for a better understanding of early flowering and improving early flowering-based breeding strategies in zucchini.

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    Comprehensive analysis of the full-length transcripts and alternative splicing involved in clubroot resistance in Chinese cabbage
    SU He-nan, YUAN Yu-xiang, YANG Shuang-juan, WEI Xiao-chun, ZHAO Yan-yan, WANG Zhi-yong, QIN Liu-yue, YANG Zhi-yuan, NIU Liu-jing, LI Lin, ZHANG Xiao-wei
    2023, 22 (11): 3284-3295.   DOI: 10.1016/j.jia.2022.09.014
    Abstract196)      PDF in ScienceDirect      

    Chinese cabbage is an economically important Brassica vegetable worldwide, and clubroot, which is caused by the soil-borne protist plant pathogen Plasmodiophora brassicae is regarded as a destructive disease to Brassica crops.  Previous studies on the gene transcripts related to Chinese cabbage resistance to clubroot mainly employed RNA-seq technology, although it cannot provide accurate transcript assembly and structural information.  In this study, PacBio RS II SMRT sequencing was used to generate full-length transcriptomes of mixed roots at 0, 2, 5, 8, 13, and 22 days after Pbrassicae infection in the clubroot-resistant line DH40R.  Overall, 39 376 high-quality isoforms and 26 270 open reading frames (ORFs) were identified from the SMRT sequencing data.  Additionally, 426 annotated long noncoding RNAs (lncRNAs), 56 transcription factor (TF) families, 1 883 genes with poly(A) sites and 1 691 alternative splicing (AS) events were identified.  Furthermore, 1 201 of the genes had at least one AS event in DH40R.  A comparison with RNA-seq data revealed six differentially expressed AS genes (one for disease resistance and five for defensive response) that are potentially involved in Pbrassicae resistance.  The results of this study provide valuable resources for basic research on clubroot resistance in Chinese cabbage.

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    Differential metabolites and their transcriptional regulation in seven major tea cultivars (Camellia sinensis) in China
    GAO Ting, HOU Bing-hao, SHAO Shu-xian, XU Meng-ting, ZHENG Yu-cheng, JIN Shan, WANG Peng-jie, YE Nai-xing
    2023, 22 (11): 3346-3363.   DOI: 10.1016/j.jia.2023.02.009
    Abstract389)      PDF in ScienceDirect      

    Various genetic and biochemical characteristics exist in tea plant cultivars, and they largely determine production suitability and tea quality.  Here, we performed transcriptomic and metabolomic analyses of young shoots of seven tea cultivars and identified major regulatory transcription factors (TFs) for the characteristic metabolites in different cultivars based on weighted gene co-expression network analysis (WGCNA).  Phenotypically, we found that ‘Tieguanyin’ (TGY) and ‘Fujian Shuixian’ (FJSX), which are suitable for oolong tea, had higher catechin contents.  The metabolites of ‘Jinxuan’ (JX) were more prominent, especially the contents of phenolic acids, flavonoids, terpenes, and tannins, which were higher than those of the other six cultivars.  Moreover, ‘Fudingdabai’ (FDDB), which is suitable for white tea, was rich in amino acids, linolenic acid, and saccharides.  At the molecular level, hydroxycinnamoyl CoA quinate hydroxycinnamoyl transferase (HCT) (CsTGY12G0001876, and CsTGY06G0003042) led to the accumulation of chlorogenic acid in TGY.  The main reason for the higher l-ascorbic acid content in FJSX was the high expression levels of L-galactono-1,4-lactone hydrogenase (GalLDH) (CsTGY13G0000389) and Myo-inositol oxygenase (MIOX) (CsTGY14G0001769, and CsTGY14G0001770), which were regulated by WRKY (CsTGY11G0001197).  Furthermore, FDDB, ‘Longjing 43’ (LJ43), ‘Shuchazao’ (SCZ)  and ‘Baihaozao’ (BHZ) had higher free fatty acid contents, among which MYB (CsTGY14G0002344) may be a hub gene for the regulation of palmitoleic acid accumulation.  More importantly, we found that the shoots of TGY were green with purple, mainly due to the accumulation of anthocyanins and the downregulation of the Mg-protoporphyrin IX nonomethyl ester cyclase (MPEC) (CsTGY10G0001989) gene that affects chlorophyll synthesis.  These results will provide a theoretical reference for tea cultivar breeding and suitability.

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    SLAF marker based QTL mapping of fruit-related traits reveals a major-effect candidate locus ff2.1 for flesh firmness in melon
    CHEN Ke-xin, DAI Dong-yang, WANG Ling, YANG Li-min, LI Dan-dan, WANG Chao, JI Peng, SHENG Yun-yan
    2023, 22 (11): 3331-3345.   DOI: 10.1016/j.jia.2023.02.014
    Abstract152)      PDF in ScienceDirect      

    Flesh firmness (FF) is an important and complex trait for melon breeders and consumers.  However, the genetic mechanism underlying FF is unclear.  Here, a soft fruit melon (P5) and a hard fruit melon (P10) were crossed to generate F2, and the FF and fruit-related traits were recorded for two years.  By performing quantitative trait locus (QTL) specific-locus amplified fragment (SLAF) (QTL-SLAF) sequencing and molecular marker-linkage analysis, 112 844 SLAF markers were identified, and 5 919 SNPs were used to construct a genetic linkage map with a total genetic distance of 1 356.49 cM.  Ten FF- and fruit-related QTLs were identified.  Consistent QTLs were detected for fruit length (FL) and fruit diameter (FD) in both years, and QTLs for single fruit weight (SFW) were detected on two separate chromosomes in both years.  For FF, the consistent major locus (ff2.1) was located in a 0.17-Mb candidate region on chromosome 2.  Using 429 F2 individuals derived from a cross between P5 and P10, we refined the ff2.1 locus to a 28.3-kb region harboring three functional genes.  These results provide not only a new candidate QTL for melon FF breeding but also a theoretical foundation for research on the mechanism underlying melon gene function.

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    Isolation and functional analysis of SrMYB1, a direct transcriptional repressor of SrUGT76G1 in Stevia rebaudiana

    ZHANG Ting, ZHANG Yong-xia, SUN Yu-ming, XU Xiao-yang, WANG Yin-jie, CHONG Xin-ran, YANG Yong-heng and YUAN Hai-yan
    2023, 22 (4): 1058-1067.   DOI: 10.1016/j.jia.2023.03.001
    Abstract206)      PDF in ScienceDirect      

    SrUGT76G1, the most well-studied diterpene glycosyltransferase in Stevia rebaudiana, is key to the biosynthesis of economically important steviol glycosides (SGs).  However, the molecular regulatory mechanism of SrUGT76G1 has rarely been explored.  In this study, we identified a MYB transcription factor, SrMYB1, using a yeast one-hybrid screening assay.  SrMYB1 belongs to the typical R2R3-type MYB protein and is specifically localized in the nucleus with strong transactivation activity.  The transcript of SrMYB1 is predominantly accumulated in flowers, but is also present at a lower level in leaves.  Yeast one-hybrid and electrophoretic mobility shift assays verified that SrMYB1 binds directly to the MYB binding sites in the F4-3 fragment (+50–(–141)) of the SrUGT76G1 promoter.  Furthermore, we found that SrMYB1 could significantly repress the expression of SrUGT76G1 in both epidermal cells of tobacco leaves and stevia callus.  Taken together, our results demonstrate that SrMYB1 is an essential upstream regulator of SrUGT76G1 and provide novel insight into the regulatory network for the SGs metabolic pathway in S. rebaudiana.

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    Core collection construction of tea plant germplasm in Anhui Province based on genetic diversity analysis using simple sequence repeat markers
    TAO Ling-ling, TING Yu-jie, CHEN Hong-rong, WEN Hui-lin, XIE Hui, LUO Ling-yao, HUANG Ke-lin, ZHU Jun-yan, LIU Sheng-rui, WEI Chao-ling
    2023, 22 (9): 2719-2728.   DOI: 10.1016/j.jia.2023.07.020
    Abstract248)      PDF in ScienceDirect      

    The tea plant [Camellia sinensis (L.) O. Kuntze] is an industrial crop in China.  The Anhui Province has a long history of tea cultivation and has a large resource of tea germplasm with abundant genetic diversity.  To reduce the cost of conservation and utilization of germplasm resources, a core collection needs to be constructed.  To this end, 573 representative tea accessions were collected from six major tea-producing areas in Anhui Province.  Based on 60 pairs of simple sequence repeat (SSR) markers, phylogenetic relationships, population structure and principal coordinate analysis (PCoA) were conducted.  Phylogenetic analysis indicated that the 573 tea individuals clustered into five groups were related to geographical location and were consistent with the results of the PCoA.  Finally, we constructed a core collection consisting of 115 tea individuals, accounting for 20% of the whole collection.  The 115 core collections were considered to have a 90.9% retention rate for the observed number of alleles (Na), and Shannon’s information index (I) of the core and whole collections were highly consistent.  Of these, 39 individuals were preserved in the Huangshan area, accounting for 33.9% of the core collection, while only 10 individuals were reserved in the Jinzhai County, accounting for 8.9% of the core set.  PCoA of the accessions in the tea plant core collection exhibited a pattern nearly identical to that of the accessions in the entire collection, further supporting the broad representation of the core germplasm in Anhui Province.  The results demonstrated that the core collection could represent the genetic diversity of the original collection.  Our present work is valuable for the high-efficiency conservation and utilization of tea plant germplasms in Anhui Province

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    Genome-wide association studies reveal the genetic basis of amino acid content variation in tea plants
    GUO Ya-fei, LI Dai-li, QIU Hai-ji, ZHANG Xiao-liang, LIU Lin, ZHAO Jing-jing, JIANG De-yuan
    2023, 22 (11): 3364-3379.   DOI: 10.1016/j.jia.2023.10.002
    Abstract170)      PDF in ScienceDirect      

    Tea is one of the most popular non-alcoholic beverages in the world, and free amino acids, especially theanine, make a major contribution to the umami taste of tea.  However, the genetic basis of the variation in amino acid content in tea plants remains largely unknown.  Here, we measured the free amino acid content in fresh leaves of 174 tea accessions over two years using a targeted metabolomics approach and obtained genotype data via RNA sequencing.  Genome-wide association studies were conducted to investigate loci affecting the content of free amino acids.  A total of 69 quantitative trait loci (–log10(P-value)>5) were identified.  Functional annotation revealed that branched-chain amino acid aminotransferase, glutamine synthetase, nitrate transporter, and glutamate decarboxylase might be important for amino acid metabolism.  Two significant loci, glutamine synthetase (Glu1, P=3.71×10–4; Arg1, P=4.61×10–5) and branched-chain amino acid aminotransferase (Val1, P=4.67×10–5; I_Leu1, P=3.56×10–6), were identified, respectively.  Based on the genotyping result, two alleles of CsGS (CsGS-L and CsGS-H) and CsBCAT (CsBCAT-L and CsBCAT-H) were selected to perform function verification.  Overexpression of CsGS-L and CsGS-H enhanced the contents of glutamate and arginine in transgenic plants, and overexpression of CsBCAT-L and CsBCAT-H promoted the accumulation of valine, isoleucine and leucine.  Enzyme activity assay uncovered that SNP1054 is important for CsGS catalyzing glutamate into glutamine.  Furthermore, CsGS-L and CsGS-H differentially regulated the accumulation of glutamine, and CsBCAT-L and CsBCAT-H differentially regulated the accumulation of branched-chain amino acids.  In summary, the findings in our study would provide new insights into the genetic basis of amino acids contents variation in tea plants and facilitate the identification of elite genes to enhance amino acids content.

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    Development and application of KASP marker for high throughput detection of the seedless trait in grapevine
    WANG Fu-qiang, BIAN Lu, QIU Peng-peng, GUO Shuo, GUO Jing-han, GUO Chen-shuo, JIANG Jian-fu, LIU Chong-huai, WANG Yong, LIU Guo-tian, WANG Yue-jin, XU Yan
    2023, 22 (11): 3269-3283.   DOI: 10.1016/j.jia.2023.10.014
    Abstract203)      PDF in ScienceDirect      

    Molecular marker-assisted selection (MAS) can significantly accelerate and improve the efficiency of the breeding process in seedless grape cultivars.  In this study, we developed the KASP_VviAGL11 and VviAGL11_410 markers based on a single nucleotide polymorphism (SNP) site (Chr18: 26889437 (A/C)) of the VviAGL11 gene, and compared them with previously reported SSR markers p3_VvAGL11 and 5U_VviAGL11 by testing 101 cultivars and 81 F1 hybrid progenies.  The results showed that both of the proposed markers obtained 100% accuracy rates in detecting allele A, which was closely associated with the seedless trait in grapes, while p3_VvAGL11 and 5U_VviAGL11 had lower accuracy rates due to their tendency to produce false positives.  After careful evaluation of the technical advantages and disadvantages associated with these markers, we concluded that KASP_VviAGL11 was superior in terms of simplicity, cost-effectiveness, efficiency, and accuracy.  Thus, we optimized the process of molecular MAS for seedless grapes, focusing on the KASP_VviAGL11 marker as a central component, to provide key technical support for the development of new seedless grape cultivars.

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    Genome wide association analysis identifies candidate genes for fruit quality and yield in Actinidia eriantha 

    Yingzhen Wang, Ying Wu, Xinlei Wang, Wangmei Ren, Qinyao Chen, Sijia Zhang, Feng Zhang, Yunzhi Lin, Junyang Yue, Yongsheng Liu
    2024, 23 (6): 1929-1939.   DOI: 10.1016/j.jia.2023.11.025
    Abstract161)      PDF in ScienceDirect      

    Quality and yield are the primary concerns in kiwifruit breeding, but research on the genetic mechanisms of fruit size, shape, and ascorbic acid (ASA) content is currently very limited, which restricts the development of kiwifruit molecular breeding.  In this study, we obtained a total of 8.88 million highly reliable single nucleotide polymorphism (SNP) markers from 140 individuals from the natural hybrid offspring of Actinidia eriantha cv. ‘White’ using whole genome resequencing technology.  A genome-wide association study was conducted on eight key agronomic traits, including single fruit weight, fruit shape, ASA content, and the number of inflorescences per branch.  A total of 59 genetic loci containing potential functional genes were located, and candidate genes related to single fruit weight, fruit length, ASA content, number of inflorescences per branch and other traits were identified within the candidate interval, such as AeWUSCHEL, AeCDK1 (cell cycle dependent kinase), AeAO1 (ascorbic oxidase) and AeCO1 (CONSTANS-like 4).  After constructing an RNAi vector for AeAO1 and injecting it into the fruit of cv. ‘Midao 31’ to interfere with the expression of the AeAO1 gene, the results showed that the activity of ascorbic oxidase in the fruit of ‘Midao 31’ significantly decreased, while the content of ASA significantly increased.  This study provides valuable insights into the genetic basis of variation in Aeriantha fruit traits, which may benefit molecular marker-assisted breeding efforts.

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    Ploidy and fruit trait variation in oil-tea Camellia: Implications for ploidy breeding
    Yanmin Li, Liangjing Yin, Xianyu He, Cenlong Hu, Ronghua Wu, Qian Long, Shixin Xiao, Deyi Yuan
    2024, 23 (8): 2662-2673.   DOI: 10.1016/j.jia.2024.03.016
    Abstract113)      PDF in ScienceDirect      
    Plant polyploidy often occurs in conjunction with higher yield and superior quality.  Therefore, obtaining polyploid germplasms is a significant part of breeding.  The oil-tea Camellia tree is an important native woody plant that produces high-quality edible oil and includes many species of Camellia with different ploidies.  However, whether higher ploidy levels in oil-tea Camellia trees are related to better traits remains unclear.  In this study, the ploidy levels of 30 different oil-tea Camellia strains in three different species in the Sect. Paracamellia were determined by flow cytometry and chromosome preparation, and the phenotypic characteristics and fatty acid compositions of the fruits were examined by field observations and laboratory analyses.  The correlations between the ploidy level of oil-tea Camellia and the main traits of the fruit were investigated.  Our results showed that 10 Camellia lanceoleosa strains were diploid, 10 Camellia meiocarpa strains were tetraploid and 10 Camellia oleifera strains were hexaploid.  Hexaploid Coleifera had larger fruit size and weight, more seeds per fruit, greater seed weight per fruit, higher oil content and greater yield per crown width than tetraploid Cmeiocarpa and diploid Clanceoleosa, but their fruit peel thickness and fresh seed rate were significantly lower, and these traits were significantly correlated with ploidy level.  In addition, in terms of fatty acid composition, hexaploid Coleifera had a higher oleic acid content than tetraploid Cmeiocarpa and diploid Clanceoleosa, but their linoleic acid, linolenic acid and arachidonic acid contents were lower.  The contents of palmitic acid, stearic acid and total unsaturated fatty acids were not significantly correlated with ploidy level.  In conclusion, certain correlations exist between the main characteristics of oil-tea Camellia fruit and the ploidy level, and increasing the ploidy level led to an increase in fruit yield with no effect on oil composition.  The discovery of variations in the main characteristics of oil-tea Camellia fruit with different ploidies will facilitate germplasm innovation and lay a foundation for ploidy breeding and mechanistic research on fruit traits.


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    Intergeneric chromosome-specific painting reveals differential chromosomal transmission from Tripidium arundinaceum in sugarcane progeny
    Fan Yu, Zehuai Yu, Jin Chai, Xikai Yu, Chen Fu, Xinwang Zhao, Hailong Chang, Jiawei Lei, Baoshan Chen, Wei Yao, Muqing Zhang, Jiayun Wu, Qinnan Wang, Zuhu Deng
    2024, 23 (11): 3751-3762.   DOI: 10.1016/j.jia.2024.08.019
    Abstract75)      PDF in ScienceDirect      
    Sugarcane has recently attracted increasing attention for its potential as a source of sugar and bioethanol, so increasing its yield is essential to ensure the sugar security and bioenergy production.  Intergeneric hybridization is a highly efficient method to produce new genetic variants of crop plants, particularly those species with high ploidy such as sugarcane (Saccharum spp.).  Tripidium arundinaceum exhibits many desirable agronomic traits, and has been widely studied to produce hybrids with improved stress tolerance and other characteristics in sugarcane breeding.  However, the genetic relationship between Tarundinaceum and Saccharum species, and the individual Tarundinaceum chromosomal compositions in sugarcane hybrids are still elusive.  Here we used whole-genome single-nucleotide polymorphisms (SNPs) to ascertain the phylogenetic relationships between these species and found that Tarundinaceum is more closely related to Saccharum than Sorghum, in contrast to the previous narrow genetic analyses using chloroplast DNA.  Additionally, oligonucleotide (oligo)-based chromosome-specific painting derived from Saccharum officinarum was able to distinctly identify the chromosomes of Tarundinaceum.  We developed the oligo-genomic in situ hybridization (GISH) system for the first time, to unveil the novel chromosome translocations and the transmission of individual Tarundinaceum chromosomes in sugarcane progeny.  Notably, we discovered that the chromosomal transmission of T. arundinaceum exhibited several different inheritance modes, including n, 2n, and over 2n in the BC1 progenies.  Such inheritance patterns may have resulted from first division restitution (FDR) or FDR+nondisjunction of a chromosome with the sister chromatids in the second meiosis division/second division restitution (FDR+NSC/SDR) model during meiosis.  These results will be of substantial benefit for the further selection of T. arundinaceum chromosomes for sugarcane genetic improvement.


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