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浙江地方特色品种温岭高橙无核潜力新种质创制及分子鉴定

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果树学报 2024年第4期41卷 590-597页

作者：张成磊师小舒陈昊谢善鹏卢鑫伍小萌刘高平郭文武解凯东台州科技职业学院·台州市园艺生物技术重点实验室浙江台州318020 华中农业大学果蔬园艺作物种质创新与利用全国重点实验室武汉430070 浙江省台州市黄岩区果树技术推广总站浙江台州318020

【目的】温岭高橙是浙江台州地方特色品种,栽培历史悠久,且具有一定的保健和药用价值,但果实有核限制了其在台州以外地区的发展,因此利用细胞工程高效育种技术对温岭高橙进行无核化改良。【方法】通过2条育种途径,一是以二倍体温岭高橙... 详细信息

【目的】温岭高橙是浙江台州地方特色品种,栽培历史悠久,且具有一定的保健和药用价值,但果实有核限制了其在台州以外地区的发展,因此利用细胞工程高效育种技术对温岭高橙进行无核化改良。【方法】通过2条育种途径,一是以二倍体温岭高橙为母本与异源四倍体为父本倍性杂交,授粉后90 d利用幼胚离体挽救培养和流式细胞仪倍性鉴定创制三倍体无核新种质;二是以雄性不育胞质杂种华柚2号为母本与温岭高橙有性杂交,通过成熟种子催芽播种,创制二倍体无核新种质。利用SSR分子标记对杂交后代进行分子鉴定。【结果】以温岭高橙为母本、异源四倍体NS和BDZNS为父本配置2个倍性杂交组合,授粉145朵花,坐果74个;胚抢救幼嫩种子1520粒,离体培养获得再生植株553株;用流式细胞仪对再生植株进行倍性鉴定,获得三倍体45株、四倍体8株;以华柚2号为母本与温岭高橙有性杂交,授粉69朵花,坐果20个,获得成熟种子312粒;通过催芽播种获得实生后代203株,倍性鉴定均为二倍体;用SSR分子标记对2个倍性杂交组合的21株三倍体和8株四倍体后代及华柚2号×温岭高橙随机选取的29株二倍体后代进行分子鉴定,显示所鉴定的三倍体及二倍体后代均含有父本特异条带,为其杂交双亲的有性后代,而倍性杂交获得的四倍体后代与母本条带完全一致,推测其为母本珠心细胞自然加倍形成的同源四倍体(双二倍体)。【结论】研究结果为温岭高橙无核新品种培育奠定了宝贵的材料基础。

关键词：柑橘无核育种倍性杂交华柚2号

维普期刊数据库博看期刊

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柑橘2个三倍体果实囊衣发育与化渣性评价

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

柑橘不同倍性种质胚性愈伤组织诱导及无病毒植株再生

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果树学报 2024年第7期41卷 1332-1341页

作者：任家勤黄雪冰杨雯惠陈昊邓家锐敖义俊解凯东郭文武伍小萌华中农业大学果蔬园艺作物种质创新与利用全国重点实验室武汉430070 城固县果业技术指导站陕西城固723200

【目的】通过离体培养败育胚珠诱导柑橘二倍体和四倍体种质的胚性愈伤组织,为柑橘多倍体研究提供成对离体材料;继续诱导胚性愈伤组织再生无毒种苗实现柑橘无核品种提纯复壮。【方法】从柑橘成熟果实中挑取败育胚珠,接种于3种愈伤诱导培... 详细信息

【目的】通过离体培养败育胚珠诱导柑橘二倍体和四倍体种质的胚性愈伤组织,为柑橘多倍体研究提供成对离体材料;继续诱导胚性愈伤组织再生无毒种苗实现柑橘无核品种提纯复壮。【方法】从柑橘成熟果实中挑取败育胚珠,接种于3种愈伤诱导培养基中,在离体培养过程中诱导并驯化胚性愈伤组织;再诱导愈伤组织分化为胚状体和不定芽,将不定芽嫁接到枳橙砧木形成完整植株;通过InDel标记和病毒检测鉴定再生植株的遗传来源和脱毒效果。【结果】离体培养诱导出13个柑橘种质的胚性愈伤组织,其中8个愈伤组织完成驯化,分别为红橘二倍体/四倍体(2x/4x)、W.默科特橘橙2x/4x、鄂柑1号椪柑2x/4x、日辉橘2x和早红脐橙。基因型影响柑橘胚性愈伤组织诱导效率,早红脐橙诱导率高达74.73%,而红橘诱导率仅6.85%;柑橘四倍体愈伤组织诱导率(0.56%~22.61%)低于对应二倍体(6.85%~37.75%)。3种不同培养基的胚性愈伤组织诱导效率不同,其中MGS培养基对温州蜜柑在内的5个品种的诱导率较高(14.13%~63.01%),MK培养基对温州蜜柑在内的4个品种的诱导率较低(21.62%~69.51%),但MK培养基对椪柑2x和红橘2x的诱导率分别为70.21%和17.31%。分别获得早红脐橙、伦晚脐橙、国庆1号温州蜜柑、大分4号温州蜜柑和兴津温州蜜柑5个无核品种的愈伤组织再生苗23、22、20、10和15株;嫁接嵌合体早红脐橙的愈伤组织再生苗经InDel标记鉴定实为罗伯逊脐橙;国庆1号温州蜜柑、伦晚脐橙和早红脐橙再生苗经PCR检测证明未感染黄脉病和衰退病病毒。【结论】柑橘胚性愈伤组织诱导率受品种基因型和培养基影响;同一品种的不同倍性种质,二倍体愈伤组织诱导率和胚状体发生率均较高;胚性愈伤组织再生植株实现了病毒完全脱除,是柑橘无核品种提纯复壮的有效手段。

关键词：柑橘胚性愈伤组织倍性种质无核品种离体再生

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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

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

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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沃柑混倍体新种质的鉴定及评价

果树学报 2024年第7期41卷 1265-1274页

作者：赵洪涛陆珍李果果解凯东郑吉祥梁增刘要鑫欧智涛梁春王举兵周双云陈东奎陈香玲广西壮族自治区农业科学院园艺研究所南宁530007 华中农业大学园艺林学学院武汉430070 广西壮族自治区水果技术指导站南宁530022 广西桂洁农业开发有限公司南宁530041

【目的】鉴定和评价沃柑变异材料,为利用提供依据。【方法】采用流式细胞仪、石蜡切片和扫描电镜方法进行倍性和叶片显微结构鉴定,比较形态特征和果实品质。【结果】变异材料为二、四混倍体(2x+4x),倍性细胞比例为2x∶4x=39.1∶60.9;混... 详细信息

【目的】鉴定和评价沃柑变异材料,为利用提供依据。【方法】采用流式细胞仪、石蜡切片和扫描电镜方法进行倍性和叶片显微结构鉴定,比较形态特征和果实品质。【结果】变异材料为二、四混倍体(2x+4x),倍性细胞比例为2x∶4x=39.1∶60.9;混倍体的叶片长、宽、周长和面积比二倍体显著增加14.75%、31.01%、19.38%和46.09%,叶形指数显著减小12.61%;两者均为两性花,花瓣数、花瓣颜色、花丝离合状态和花柱状态相同,花瓣长度和宽度均显著大于二倍体;种子均为单胚类型,单粒质量、横径和纵径较二倍体分别显著增加30.77%、8.12%和26.64%;单果质量、纵径、横径、果形指数、果皮厚度和果实硬度比二倍体分别显著增加29.67%、13.38%、8.57%、3.85%、38.31%和16.37%,单果种子数显著减少61.68%;可溶性固形物和可滴定酸含量无显著差异,可食率显著降低,出汁率高,维生素C含量显著增加39.70%;叶片、下表皮、栅栏组织和海绵组织厚度较二倍体分别显著增加19.75%、10.10%、9.38%和43.92%,气孔长度和密度分别显著减小7.36%和15.76%,气孔宽度显著增加5.09%。【结论】鉴定获得了沃柑二、四混倍体新种质,与二倍体在形态特征、果实品质及叶片显微结构等方面存在较大差异。

关键词：沃柑二倍体混倍体形态特征果实品质显微结构

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Mass extinctions, their causes and consequences: an interview with Douglas *** and Shuzhong Shen

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National Science Review 2024年第1期11卷 66-70页

作者： Shucheng xie State Key Laboratory of Biogeology and Environmental Geology China University of Geosciences at Wuhan

‘Mass extinctions’ have been a hot topic for several decades. What triggers a mass extinction? How does a mass extinction impact the evolution of life? How does our ecosystem recover after a mass extinction? These questions attracted the interest of both scientists and the public *** spoke to two renowned researchers in the field of mass extinctions: Prof. Douglas H. Erwin from the National Museum of Natural History of the USA, and Prof. Shuzhong Shen from Nanjing University, China.

关键词： Stratigraphy

Turbulence Statistics of Thermo-Buoyancy Supercritical Fuel Flow in a Regenerative Cooling Channel

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Journal of Thermal Science 2024年第1期33卷 126-137页

作者： SUN Feng xie Gongnan School of Marine Science and Technology Northwestern Polytechnical UniversityXi'an 710072China Department of Thermal Engineering Tsinghua UniversityBeijing 100084China

Active regenerative cooling with supercritical hydrocarbon fuel is considered as the most promising thermal protection *** existence of buoyancy force would lead to strongly anisotropic flow and thermal transport *** is closely related to the cooling performance of the *** elucidate the mechanisms of turbulent transport,the large eddy simulation(LES) was performed to assess turbulence statistics within different turbulence *** results indicated that the buoyancy and inertial force together dominated the change of turbulent ***,the spatial thermal buoyancy effect significantly suppressed the vertical velocity *** is due to the laminar motion caused by the buoyancy force,thereby weakening the thermal *** the statistics of velocity fluctuation,it was found that the buoyancy force and inertial force greatly weaken the vertical and streamwise velocity fluctuation,*** the statistics of thermal transport,the results pointed out that the near-wall heat transport characteristics need to be paid more *** thickness of the temperature mixing boundary layer led to the attenuation of vertical heat flux,which inhibited vertical temperature diffusion and predisposed to extreme conditions of heat transfer *** results can enhance the academic understanding of the heat transfer mechanism of supercritical fluids,and give guidance for further applications of thermal protection.

关键词： active regenerative cooling large eddy simulation(LES) buoyancy force turbulence statistics