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Peculiarity of transcriptional and H3K27me3 dynamics during peach bud dormancy

Horticultural Plant Journal 2024年第1期10卷 38-50页

作者： Yalin Zhao Yong Li Ke Cao Weichao Fang Changwen Chen Xinwei Wang Jinlong Wu wenwu guo Lirong Wang The Key Laboratory of Biology and Genetic Improvement of Horticultural Crops(Fruit Tree Breeding Technology) Ministry of AgricultureZhengzhou Fruit Research InstituteChinese Academy of Agricultural SciencesZhengzhouHenan 450009China Western Research Institute Chinese Academy of Agricultural SciencesChangjiXinjiang 831100China College of Horticulture and Forestry Science Huazhong Agricultural UniversityWuhanHubei 430070China

Bud dormancy facilitates the survival of meristems under harsh environmental *** elucidate how molecular responses to chilling accumulation controlling dormancy in peach buds,chromatin immunoprecipitation sequencing to identify the H3K27me3 modifications and RNA sequencing of two peach cultivars with pronounced differences in chilling requirement were carried out,the results showed that genes associated with abscisic acid and gibberellic acid signal pathways play key roles in dormancy *** results demonstrated that peach flower bud differentiation occurred continuously in both cultivars during chilling accumulation,which was correlated with the transcript abundance of key genes involved in phytohormone metabolism and flower bud development under adverse *** more increased strength in high chillingrequirement cultivar along with the chilling accumulation at the genome-wide *** function of the dormancy-associated MADS-box gene PpDAM6 was identified,which is involved in leaf bud break in peach and flower development in transgenic Nicotiana tabacum(NC89).In addition,PpDAM6 was positively regulated by PpCBF,and the genes of putative dormancy-related and associated with metabolic pathways were *** together,these results constituted a theoretical basis for elucidating the regulation of peach bud dormancy transition.

关键词： Peach bud Dormancy RNA-seq H3K27me3 Chilling requirement

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大果少核椪柑新品种华柑3号的选育

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果树学报 2024年第3期41卷 543-546页

作者：解凯东陈昊伍小萌谢宗周郭文武华中农业大学园艺林学学院·果蔬园艺作物种质创新与利用全国重点实验室武汉430070

华柑3号是通过鄂柑1号椪柑珠心胚实生苗倍性变异而培育的四倍体大果少核椪柑新品种。果实扁圆形,果实大,平均横径74.08 mm,纵径62.63 mm,平均单果质量179.94 g,平均果皮厚度3.45 mm。果肉橙黄色,肉质脆嫩,酸甜爽口,风味浓,可溶性固形物... 详细信息

华柑3号是通过鄂柑1号椪柑珠心胚实生苗倍性变异而培育的四倍体大果少核椪柑新品种。果实扁圆形,果实大,平均横径74.08 mm,纵径62.63 mm,平均单果质量179.94 g,平均果皮厚度3.45 mm。果肉橙黄色,肉质脆嫩,酸甜爽口,风味浓,可溶性固形物含量(w,后同)为14.49%,可滴定酸含量为1.16%,可食率为71.6%,化渣性好;种子少,平均2.7粒·果^(-1);树势中等偏强,树姿较直立,结果母枝以春梢和秋梢为主;在湖北武汉地区,12月中下旬果实成熟,可挂树至翌年1月;丰产性中等,按3.0 m×4.0 m密度定植的5年生树,每666.7 m^(2)产量可达1500 kg;适宜在年均温度16℃左右,冬季无霜冻、绝对最低温度在-5℃以上的区域种植。

关键词：柑橘新品种华柑3号多倍体无核育种

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DR5 is a Suitable System for Studying the Auxin Response in the Poncirus trifoliata-Xanthomonas axonopodis *** Interaction

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Horticultural Plant Journal 2020年第5期6卷 277-283页

作者： Xuan Xiao Xiaoyan Li Chunli Chen wenwu guo College of Life Science Gannan Normal UniversityGanzhouJiangxi 341000China Key Laboratory of Horticultural Plant Biology(Ministry of Education) College of Horticulture and Forestry SciencesHuazhong Agricultural UniversityWuhan 430070China

The local auxin distribution characteristics in the roots,stems,and leaves of stably transformed plantlets of trifoliate orange(Poncirus trifoliata)with auxin reporter system DR5::GUS-YFP were elucidated in this *** auxin response maxima could be observed in the apex of the root tip,primary phloem of the tender stem,and the margin of the young leaves according to the activity of theβ-glucuronidase(GUS)reporter gene triggered by the auxin responsive DR5 *** responses in the apex of the root tips increased when treated with synthetic auxin 1-naphthylacetic acid(NAA),but decreased when treated with the auxin polar transportation inhibitor 2,3,5-triiodobenzoic acid(TIBA).These results indicated that the DR5 reporter system worked in *** for auxin distribution and response *** orange is highly susceptible to citrus canker *** accumulation was observed visually in the invasion sites of the detached leaves inoculated with Xanthomonas axonopodis ***(Xac)by GUS staining;the upregulated expression of the YFP,GH3.1,GH3.9,and SAUR genes assessed by quantitative real-time PCR(qRT-PCR)also identified auxin accumulation in the inoculated tissues following Xac ***,these findings indicated that the plantlets of *** engineered with the auxin reporter gene provided a promising system for studying auxin responses during Xac infection.

关键词： Poncirus trifoliata DR5 auxin Xanthomonas axonopodis pv.citri

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Mitochondrial Genome of Callus Protoplast Has a Role in Mesophyll Protoplast Regeneration in Citrus: Evidence From Transgenic GFP Somatic Homo-Fusion

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Horticultural Plant Journal 2017年第5期3卷 177-182页

作者： CAI Xiaodong FU Jing guo wenwu Key Laboratory of Horticultural Plant Biology (Ministry of Education) College of Horticulture and Forestry Sciences Huazhong Agricultural University College of Horticulture and Gardening Yangtze University

Protoplast fusion has great potential in citrus improvement. Although citrus mesophyll protoplasts usually cannot divide and regenerate,symmetric protoplast fusion of embryogenic callus protoplast + mesophyll protoplast sometimes results in the regeneration of mesophyllparent-type cybrids. It suggested that mitochondrial DNA(mt DNA) from protoplasts of embryogenic callus parent plays an important role in stimulating division and regeneration of mesophyll protoplasts. Herein, somatic fusion was conducted via electrofusion between callus protoplasts isolated from Valencia orange [Citrus sinensis(L.) Osbeck] cell suspension cultures and transgenic GFP-tagged mesophyll protoplasts from the same genotype, i.e. transgenic Valencia orange plants containing the green fluorescent protein(GFP) gene, in an effort to elucidate whether mt DNA of callus line could stimulate the division and regeneration of mesophyll protoplasts from the same genotype. Two embryoids and one plantlet with GFP expression were successfully obtained and subsequent ploidy analysis by flow cytometry indicated that they were all diploids. The regenerated diploid embryoids and plantlet with GFP expression could be considered as ‘cybrids' with mt DNA from the callus protoplasts of Valencia orange. The result indicated that citrus mesophyll-parent-type cybrid regeneration needed the stimulation of mt DNA from protoplasts of embryogenic callus parent regardless of their origin either from another genotype or the same genotype as the mesophyll parent.

关键词： citrus cybrid mitochondrial DNA(mtDNA) callus protoplast mesophyll somatic hybridization

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柑橘2个三倍体果实囊衣发育与化渣性评价

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园艺学报 2024年第6期51卷 1179-1188页

作者：曹惠祥罗鑫刘婉荣管书萍王婷婷伍小萌郭文武解凯东华中农业大学园艺林学学院果蔬园艺作物种质创新与利用全国重点实验室武汉430070 襄阳市农业科学院湖北襄阳441100

探讨柑橘倍性杂交三倍体后代及其二倍体母本果实化渣性差异及原因,为柑橘三倍体果实化渣相关研究奠定数据基础。以本地早橘和红橘为对照,分别对以其为母本与四倍体倍性杂交获得的2个三倍体有性后代果实囊衣发育过程中的厚度变化和组分... 详细信息

探讨柑橘倍性杂交三倍体后代及其二倍体母本果实化渣性差异及原因,为柑橘三倍体果实化渣相关研究奠定数据基础。以本地早橘和红橘为对照,分别对以其为母本与四倍体倍性杂交获得的2个三倍体有性后代果实囊衣发育过程中的厚度变化和组分含量进行分析,探究囊衣结构及其成分对化渣性的影响。结果表明:2个三倍体后代果实囊衣厚度、细胞层数、硬度和剪切力在不同发育时期均显著大于其二倍体母本;与二倍体母本相比,2个三倍体后代囊衣细胞壁更厚,微纤丝密度更大,结构更紧密;囊衣的纤维素、半纤维素、水溶性果胶和原果胶含量均显著高于其二倍体母本,木质素含量则在部分发育时期显著低于二倍体母本。综上所述,果实囊衣厚,纤维素、半纤维素和果胶含量高可能是柑橘部分三倍体化渣性较差的主要原因,而这些指标可用于三倍体化渣性的评价和筛选。

关键词：柑橘三倍体囊衣化渣性纤维素含量

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An efficient transient gene expression system for protein subcellular localization assay and genome editing in citrus protoplasts

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Horticultural Plant Journal 2023年第3期9卷 425-436页

作者： Wenhui Yang Jiaqin Ren Wanrong Liu Dan Liu Kaidong Xie Fei Zhang Pengwei Wang wenwu guo Xiaomeng Wu National Key Laboratory for Germplasm Innovation&Utilization of Horticultural Crops College of Horticulture and Forestry SciencesHuazhong Agricultural UniversityWuhanHubei 430070China Hubei Hongshan Laboratory WuhanHubei 430070China

Protoplast has been widely used in biotechnologies to circumvent the breeding obstacles in citrus, including long juvenility, polyembryony, and male/female sterility. The protoplast-based transient gene expression system is a powerful tool for gene functional characterization and CRISPR/Cas9 genome editing in higher plants, but it has not been widely used in citrus. In this study, the polyethylene glycol(PEG)-mediated method was optimized for citrus callus protoplast transfection, with an improved transfection efficiency of 68.4%. Consequently, the efficiency of protein subcellular localization assay was increased to 65.8%, through transient expression of the target gene in protoplasts that stably express the fluorescent organelle marker protein. The gene editing frequencies in citrus callus protoplasts reached 14.2% after transient expression of CRISPR/Cas9 constructs. We demonstrated that the intronic polycistronic tRNAgRNA(inPTG) genome editing construct was functional in both the protoplast transient expression system and epicotyl stable transformation system in citrus. With this optimized protoplast transient expression system, we improved the efficiency of protein subcellular localization assay and developed the genome editing system in callus protoplasts, which provides an approach for prompt test of CRISPR vectors.

关键词： Citrus Callus protoplast Transient transfection Subcellular localization Genome editing

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与呼吸跃变型果实共贮藏改善柑橘果实色泽和品质

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园艺学报 2024年第3期51卷 601-615页

作者：孙权何政辰叶俊丽魏冉冉尹映紫柴利军谢宗周徐强徐娟郭文武程运江邓秀新华中农业大学园艺植物生物学教育部重点实验室国家柑橘育种中心武汉430070

选择9种常见园艺果实分别与纽荷尔脐橙果实进行采后共贮藏,结果表明:与呼吸跃变型果实(香梨、猕猴桃、番茄、香蕉)共贮藏(storage with climacteric fruits,SWCF)处理可以显著促进纽荷尔脐橙果皮转红,其中香梨的效果最佳。高效液相色谱... 详细信息

选择9种常见园艺果实分别与纽荷尔脐橙果实进行采后共贮藏,结果表明:与呼吸跃变型果实(香梨、猕猴桃、番茄、香蕉)共贮藏(storage with climacteric fruits,SWCF)处理可以显著促进纽荷尔脐橙果皮转红,其中香梨的效果最佳。高效液相色谱分析显示,SWCF处理促进纽荷尔脐橙果皮总类胡萝卜素、β–柠乌素、β–隐黄质和紫黄质含量增加3倍左右。其中,β–柠乌素的积累是SWCF处理促进柑橘果皮转红的主要原因。实时荧光定量PCR结果显示,SWCF处理显著激活了类胡萝卜素代谢基因的表达,特别是类胡萝卜素裂解双加氧酶4b基因(Carotenoid cleavage dioxygenase 4b,Cs CCD4b)最大被激活了约20倍,这是SWCF处理促进β–柠乌素积累的直接原因。进一步研究发现,SWCF处理改善果皮色泽对各品种柑橘具有普遍性。此外,与乙烯处理相比,SWCF处理显著降低了柑橘果蒂衰老率、失重率以及果皮蜡质的降解,增加了果实挥发性物质的含量。SWCF处理为绿色高效改善柑橘果实外观品质提供了新思路。

关键词：柑橘呼吸跃变型果实生物处理果皮色泽果实品质采后贮藏处理

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柑橘2个三倍体有性后代群体果实品质性状的遗传特点

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果树学报 2024年第3期41卷 369-378页

作者：管书萍王婷婷周阳广朱虹娴伍小萌龙春瑞高俊燕郭文武解凯东华中农业大学园艺林学学院·果蔬园艺作物种质创新与利用全国重点实验室武汉430070 云南省农业科学院热带亚热带经济作物研究所云南保山678000

【目的】研究柑橘三倍体有性后代果实品质性状的遗传特点,为倍性杂交创制无核三倍体新种质科学选配亲本提供理论依据。【方法】以柑橘二倍体品种秋辉橘、清见橘橙为母本,异源四倍体体细胞杂种橘柚+甜橙(NS)、甜橙+红橘(SD)为父本倍性杂... 详细信息

【目的】研究柑橘三倍体有性后代果实品质性状的遗传特点,为倍性杂交创制无核三倍体新种质科学选配亲本提供理论依据。【方法】以柑橘二倍体品种秋辉橘、清见橘橙为母本,异源四倍体体细胞杂种橘柚+甜橙(NS)、甜橙+红橘(SD)为父本倍性杂交获得的2个三倍体有性后代群体成熟果实为材料,通过测定其果实横纵径、单果质量、果皮厚度、可溶性固形物和可滴定酸含量等性状,探究柑橘三倍体有性后代果实品质性状的遗传特点。【结果】三倍体后代单果质量和果实横径平均值均介于双亲之间,呈趋中变异;果实纵径变化趋势在2个组合间有差异,秋辉橘×NS组合后代呈趋大变异,而清见橘橙×SD组合则呈趋中变异;果皮厚度平均值均大于亲中值且多数高于高值亲本,存在一定程度的超高亲本遗传现象;2个组合三倍体有性后代果实可溶性固形物含量平均低低亲比率分别为74.4%、66.88%,呈趋小变异;而2个三倍体有性后代群体果实可滴定酸含量平均值均低于亲中值,且不同年份间酸含量介于亲本间和低于低值亲本的后代比例较高。【结论】推测柑橘三倍体有性后代果实单果质量、果实横径、纵径、果形指数、果皮厚度和可溶性固形物含量性状是由多基因控制的数量性状,可滴定酸含量性状可能存在主效控制基因。

关键词：柑橘三倍体倍性育种果实性状遗传倾向

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Overexpression of miR390b promotes stem elongation and height growth in Populus

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Horticulture Research 2023年第2期10卷 65-77页

作者： Qiaofang Shi Dongdong Tian Jieyu Wang Aoli Chen Yuqing Miao Yiming Chen Jun Li Xiaomeng Wu Bo Zheng wenwu guo Xueping Shi Key Laboratory of Horticultural Plant Biology of Ministry of Education College of Horticulture and Forestry SciencesHuazhong Agricultural UniversityWuhan 430070China Poplar Research Center Huazhong Agricultural UniversityWuhan 430070China Tobacco Research Institute Chinese Academy of Agricultural SciencesQingdao 266101China

MicroRNA390(miR390)is involved in plant growth and development by down-regulating the expression of the downstream genes trans-acting short interfering RNA3(TAS3)and AUXIN RESPONSE FACTORs(ARFs).There is a scarcity of research on the involvement of the miR390-TAS3-ARFs pathway in the stem development of ***,differentially expressedmiRNAs during poplar stem development were screened by small RNA sequencing analysis,and a novel function of miR390b in stem development was *** of miR390b(OE-miR390b)resulted in a large increase in the number of xylem fiber cells and a slight decrease in the cell length at the longitudinal *** increases in stem elongation and plant height were observed in the OE-miR390b *** to transcriptome sequencing results and transient co-expression analysis,TAS3.1 and TAS3.2 were identified as the target genes of miR390 in poplar and were negatively regulated by miR390 in the *** transcription levels of ARF3.2 and ARF4 were significantly repressed in OE-miR390b plants and strongly negatively correlatedwith the number of xylem fiber cells along the longitudinal *** findings indicate that the conserved miR390-TAS3-ARFs pathway in poplar is involved in stem elongation and plant height growth.

关键词： analysis transient stem

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UNREDUCED MEGAGAMETOPHYTE FORMATION VIA SECOND DIVISION RESTITUTION CONTRIBUTES TO TETRAPLOID PRODUCTION IN INTERPLOIDY CROSSES WITH/ORAH/MANDARIN(CITRUS RETICULATA)

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Frontiers of Agricultural Science and Engineering 2021年第2期8卷 302-313页

作者： Qiangming XIA Wei WANG Kaidong XIE Xiaomeng WU Xiuxin DENG Jude WGROSSER wenwu guo Key Laboratory of Horticultural Plant Biology(Ministry of Education) College of Horticulture and Forestry SciencesHuazhong Agricultural UniversityWuhan 430070China Citrus Research and Education Center University of Florida/IFASLake AlfredUSA.

Seedless fruits are desirable in the citrus fresh fruit *** production via diploid x tetraploid interploidy crosses is thought to be the most efficient and widely-used strategy for the breeding of seedless ***'Orah'mandarin has desirable organoleptic qualities,seeds in the fruits weaken its market *** produce new seedless cultivars that are similar to'Orah'mandarin,we performed three 2x x 4x crosses using'Orah'mandarin as the seed parent to regenerate triploid plantlets.A total of 182 triploid and 36 tetraploid plantlets were *** analyzing their genetic origins using nine novel single nucleotide polymorphism(SNP)markers,all of the triploids and tetraploids derived from these three crosses were proven to be ***,we demonstrated that 2n megagametophyte formation in'Orah'mandarin result in tetraploid production in these three interploidy *** tetraploid plantlets were genotyped using eight pericentromeric SNP markers and nine centromere distal SNP *** on the genotypes of the 2n megagametophytes,the parental heterozygosity rates in 16 SNP loci and all 2n megagametophytes were less than 50%,indicating that second division restitution was the mechanism underlying 2n megagametophyte formation at both the population and individual *** triploid hybrids enrich the germplasm available for seedless ***,the tetraploid hybrids are valuable as parents for ploidy breeding for the production of seedless citrus fruits.

关键词： Citrus 2n gamete interploidy hybridization pericentromeric SNP marker second division restitution

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