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Transcription factor CsTT8 promotes fruit coloration by positively regulating the methylerythritol 4-phosphate pathway and carotenoid biosynthesis pathway in citrus (Citrus spp.)

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Horticulture Research 2023年第11期10卷 71-81页

作者： Quan Sun Zhengchen He Ranran Wei Yingzi Yin Junli Ye Lijun Chai zongzhou xie Wenwu Guo Juan Xu Yunjiang Cheng Qiang Xu Xiuxin Deng National Key Laboratory for Germplasm Innovation and Utilization of Horticultural Crops Huazhong Agricultural UniversityWuhan 430070China National Research Center for Apple Engineering and Technology Shandong Agricultural UniversityTaianShandong 271018China Hubei Hongshan Laboratory Wuhan Hubei 430070China

Carotenoids directly influence citrus fruit color and nutritional value,which is critical to consumer acceptance.Elucidating the potential molecular mechanism underlying carotenoid metabolism is of great importance for improving fruit quality.Despite the well-established carotenoid biosynthetic pathways,the molecular regulatory mechanism underlying carotenoid metabolism remains poorly understood.Our previous studies have reported that the Myc-type basic helix-loop-helix(bHLH)transcription factor(TF)regulates citrus proanthocyanidin biosynthesis.Transgenic analyses further showed that overexpression of CsTT8 could significantly promote carotenoid accumulation in transgenic citrus calli,but its regulatory mechanism is still unclear.In the present study,we found that overexpression of CsTT8 enhances carotenoid content in citrus fruit and calli by increasing the expression of CsDXR,CsHDS,CsHDR,CsPDS,CsLCYE,CsZEP,and CsNCED2,which was accompanied by changes in the contents of abscisic acid and gibberellin.The in vitro and in vivo assays indicated that CsTT8 directly bound to the promoters of CsDXR,CsHDS,and CsHDR,the keymetabolic enzymes of the methylerythritol 4-phosphate(MEP)pathway,thus providing precursors for carotenoid biosynthesis and transcriptionally activating the expression of these three genes.In addition,CsTT8 activated the promoters of four key carotenoid biosynthesis pathway genes,CsPDS,CsLCYE,CsZEP,and CsNCED2,directly promoting carotenoid biosynthesis.This study reveals a novel network of carotenoid metabolism regulated by CsTT8.Our findings will contribute to manipulating carotenoid metabolic engineering to improve the quality of citrus fruit and other crops.

关键词： citrus Citrus metabolism

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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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Citrus β-carotene hydroxylase 2 (BCH2) participates in xanthophyll synthesis by catalyzing the hydroxylation of β-carotene and compensates for BCH1 in citrus carotenoid metabolism

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Horticulture Research 2023年第3期10卷 45-56页

作者： Yingzi Zhang Jiajing Jin Shenchao Zhu Quan Sun Yin Zhang zongzhou xie Junli Ye Xiuxin Deng Key Laboratory of Horticultural Plant Biology(Ministry of Education) Huazhong Agricultural UniversityWuhan430070China

Abstract As an essential horticultural crop,Citrus has carotenoid diversity,which affects its aesthetic and nutritional values.β,β-Xanthophylls are the primary carotenoids accumulated in citrus fruits,and non-heme di-iron carotene hydroxylase(BCH)enzymes are mainly responsible forβ,β-xanthophyll synthesis.Previous studies have focused on the hydroxylation of BCH1,but the role of its paralogous gene in citrus,BCH2,remains largely unknown.In this study,we revealed theβ-hydroxylation activity of citrus BCH2(CsBCH2)for the first time through the functional complementation assay using Escherichia coli,although CsBCH2 exhibited a lower activity in hydroxylatingβ-carotene intoβ-cryptoxanthin than citrus BCH1(CsBCH1).Our results showed that overexpression of CsBCH2 in citrus callus increased xanthophyll proportion and plastoglobule size with feedback regulation of carotenogenic gene expression.This study revealed the distinct expression patterns and functional characteristics of two paralogous genes,CsBCH1 and CsBCH2,and illustrated the backup compensatory role of CsBCH2 for CsBCH1 in citrus xanthophyll biosynthesis.The independent function of CsBCH2 and its cooperative function with CsBCH1 inβ-cryptoxanthin biosynthesis suggested the potential of CsBCH2 to be employed for expanding the synthetic biology toolkit in carotenoid engineering。

关键词： citrus Citrus metabolism

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Natural Variation in CCD4 Promoter Underpins Species-Specific Evolution of Red Coloration in Citrus Peel

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Molecular Plant 2019年第9期12卷 1294-1307页

作者： Xiongjie Zheng Kaijie Zhu Quan Sun Weiyi Zhang Xia Wang Hongbo Cao Meilian Tan zongzhou xie Yunliu Zeng Junli Ye Lijun Chai Qiang Xu Zhiyong Pan Shunyuan Xiao Paul D.Fraser Xiuxin Deng Key Laboratory of Horticultural Plant Biology(Ministry of Education) Huazhong Agricultural University.WuhanChina School of Biological Sciences.Royal Holloway University of LondonEghamSurrey TW20 OEXUK Department of Plant Science and Landscape Architecture Institute for Bioscience and Biotechnology ResearchUniversity of MarylandRockvilleMDUSA Present address:College of Horticulture Agricultural University of Hebei.Baoding Hebei 071000China Present address:Key Laboratory of Biology and Genetic Improvement of Oil Crops Ministry of Agriculture.Oil Crops Research Institute of the Chinese Academy of Agricultural SciencesWuhan 430070China

Carotenoids and apocarotenoids act as phytohormones and volatile precursors that influence plant development and confer aesthetic and nutritional value critical to consumer preference.Citrus fruits display considerable natural variation in carotenoid and apocarotenoid pigments.In this study,using an integrated genetic approach we revealed that a 5;c/s-regulatory change at CCD4b encoding CAROTENOID CLEAVAGE DIOXYGENASE 4b is a major genetic determinant of natural variation in C3 0 apocarotenoids responsible for red coloration of citrus peel.Functional analyses demonstrated that in addition the known role in synthesizing 3-citraurin,CCD4b is also responsible for the production of another important C3 0 apocarotenoid pigment,p-citraurinene.Furthermore,analyses of the CCD4b promoter and transcripts from various citrus germplasm accessions established a tight correlation between the presence of a putative 5'c/s-regulatory enhancer within an MITE transposon and the enhanced allelic expression of CCD4b in C3 0 apocarotenoid-rich red-peeled accessions.Phylogenetic analysis provided further evidence that functional diversification of CCD4b and naturally occurring variation of the CCD4b promoter resulted in the stepwise evolution of red peels in mandarins and their hybrids.Taken together,our findings provide new insights into the genetic and evolutionary basis of apocarotenoid diversity in plants,and would facilitate breeding efforts that aim to improve the nutritional and aesthetic value of citrus and perhaps other fruit crops.

关键词： citrus apocarotenoid natural variation carotenoid cleavage dioxygenase promoter transposon

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NEW INSIGHTS INTO THE PHYLOGENY AND SPECIATION OF KUMQUAT (FORTUNELLA SPP.) BASED ON CHLOROPLAST SNP, NUCLEAR SSR AND WHOLE-GENOME SEQUENCING

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Frontiers of Agricultural Science and Engineering 2022年第4期9卷 627-641页

作者： Chenqiao ZHU Peng CHEN Junli YE Hang LI Yue HUANG Xiaoming YANG Chuanwu CHEN Chenglei ZHANG Yuantao XU Xiaoli WANG Xiang YAN Guangzhou DENG Xiaolin JIANG Nan WANG Hongxing WANG Quan SUN Yun LIU Di FENG Min YU xietian SONG zongzhou xie Yunliu ZENG Lijun CHAI Qiang XU Chongling DENG Yunjiang CHENG Xiuxin DENG Key Laboratory of Horticultural Plant Biology(Ministry of Education) Huazhong Agricultural UniversityWuhan 430070China Fruit Industry Technology Guidance Stand of Ganzhou Ganzhou 341000China Guangxi Academy of Specialty Crops/Guangxi Citrus Breeding and Cultivation Research Center of Engineering Technology Guilin 541004China Ganzhou Citrus Research Institute Ganzhou 341000China

Kumquat(Fortunella spp.)is a fruit and ornamental crop worldwide due to the palatable taste and high ornamental value of its fruit.Although Fortunella is classified into the economically important true citrus fruit tree group together with Citrus and Poncirus,few studies have been focused on its evolutionary scenario.In this study,analysis of five chloroplast loci and 47 nuclear microsatellites(nSSR)loci from 38 kumquat and 10 citrus accessions revealed the independent phylogeny of Fortunella among citrus taxa,and that Fortunella mainly comprises two populations:CUL,cultivated Fortunella spp.(F.margarita,F.crassifolia and F.japonica);and HK,wild Hong Kong kumquat(Fortunella hindsii).Genomic analysis based on whole-genome SNPs indicated that the allele frequency of both pupations deviated from the neutral selection model,suggesting directional selection was a force driving their evolutions.CUL exhibited lower genomic diversity and higher linkage strength than HK,suggesting artificial selection involved in its origin.A high level of genetic differentiation(Fst=0.364)was detected and obviously asynchronous demographic changes were observed between CUL and HK.Based on these results,a new hypothesis for the speciation of Fortunella is proposed.

关键词： Citrus Fortunella kumquat phylogenetics

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Genome of Wild Mandarin and Domestication History of Mandarin

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Molecular Plant 2018年第8期11卷 1024-1037页

作者： Lun Wang Fa He Yue Huang Jiaxian He Shuizhi Yang Jiwu Zeng Chongling Deng Xiaolin Jiang Yiwen Fang Shaohua Wen Rangwei Xu Huiwen Yu Xiaoming Yang Guangyan Zhong Chuanwu Chen Xiang Yan Changfu Zhou Hongyan Zhang zongzhou xie Robert M. Larkin Xiuxin Deng Qiang Xu Key Laboratory of Horticultural Plant Biotogy (Ministry of Education) Huazhong Agricultural University Wuhan 430070 P.R. China Horticulture institute Hunan Academy of Agricultural Sciences Changsha 410125 P.R. China Fruit Tree Research Institute Guangdong Academy of Agricultural Sciences Guangzhou 510640 P.R. China Guangxi Key Laboratory of Citrus Biology Guangxi Academy of Specialty Crops Guilin 541000 P.R. China Institute of Citrus Science Research of Ganzheu City Ganzhou 341000 P.R. China

Mandarin （Citrus reticulata） is one of the most important citrus crops worldwide. Its domestication is believed to have occurred in South China, which has been one of the centers of mandarin cultivation for four millennia. We collected natural wild populations of mandarin around the Nanling region and cultivated landraces in the vicinity. We found that the citric acid level was dramatically reduced in cultivated mandarins. To understand genetic basis of mandarin domestication, we de novo assembled a draft genome of wild mandarin and analyzed a set of 104 citrus genomes. We found that the Mangshan mandarin is a primitive type and that two independent domestication events have occurred, resulting in two groups of cultivated mandarins （MD1 and MD2） in the North and South Nanling Mountains, respectively. Two bottlenecks and two expansions of effective population size were identified for the MD1 group of cultivated mandarins. However, in the MD2 group there was a long and continuous decrease in the population size. MD1 and MD2 mandarins showed different patterns of interspecific introgression from cultivated pummelo species. We identified a region of high divergence in an aconitate hydratase （ACO） gene involved in the regulation of citrate content, which was possibly under selection during the domestication of mandarin. This study provides concrete genetic evidence for the geographical origin of extant wild mandarin populations and sheds light on the domestication and evolutionary history of mandarin.

关键词： Citrus Citric acid Domestication Genome Wild mandarin

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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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无核柑橘新品种华柑4号的选育

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果树学报 2022年第3期39卷 495-498页

作者：吴巨勋张雅菁伊华林谢宗周邓秀新华中农业大学园艺林学学院·园艺植物生物学教育部重点实验室武汉430070

华柑4号是从靖安椪柑(Citrus reticulata Blanco.)芽变中选育出来的无核柑橘新品种。果实扁圆形,果形指数0.80,平均果皮厚度2.0 mm,平均单果质量124.81 g,成熟后果皮黄橙色。可溶性固形物含量(w,后同)为13.2%,可滴定酸含量1.17%,维生素... 详细信息

华柑4号是从靖安椪柑(Citrus reticulata Blanco.)芽变中选育出来的无核柑橘新品种。果实扁圆形,果形指数0.80,平均果皮厚度2.0 mm,平均单果质量124.81 g,成熟后果皮黄橙色。可溶性固形物含量(w,后同)为13.2%,可滴定酸含量1.17%,维生素C含量为30.77 mg·100 g^(-1),平均单果种子数为0.19粒,混栽或单独栽培均接近无籽;品质上。树势强,结果母枝主要以春梢和早秋梢为主,花芽分化能力强,花量中等,自然授粉坐果率5%~8%,丰产稳产。在湖北武汉地区,12月中下旬成熟。

关键词：柑橘新品种无核椪柑芽变

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采收时期和授粉对玉皇柚品质的影响

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华中农业大学学报 2022年第4期41卷 210-216页

作者：狄明慧张红艳李登朝谭碧玲谢宗周华中农业大学园艺林学学院/园艺植物生物学教育部重点实验室武汉430070 湖北省建始县林业技术推广站建始445300 湖北省建始煜立农业发展有限公司建始445300

以湖北省建始县的地方柑橘品种玉皇柚为试材,比较不同采收时期果实外观和内在品质差异,确定最佳采收期;并比较花后240 d无籽类型和有籽类型果实外观及风味物质的差异,以期为玉皇柚的采收以及生产上无籽和有籽的取舍提供理论依据。结果显... 详细信息

以湖北省建始县的地方柑橘品种玉皇柚为试材,比较不同采收时期果实外观和内在品质差异,确定最佳采收期;并比较花后240 d无籽类型和有籽类型果实外观及风味物质的差异,以期为玉皇柚的采收以及生产上无籽和有籽的取舍提供理论依据。结果显示:花后240 d时采收的果实果皮着色程度和可溶性固形物含量最高,有籽果实可滴定酸含量最低,固酸比最高为14.62,此时采收的果实外观和风味品质最佳,花后222 d时采收品质次之;而无籽果花后222 d时采收最佳,最高固酸比为14.55。花后240 d有籽果Vc含量(有籽类型35.98 mg/100 g,无籽类型31.77 mg/100 g)及以大根香叶烯D为主的挥发性物质含量也更高。结果表明,适时采收和授粉可以优化玉皇柚的果实品质,可通过科学采收和提高授粉率来提升果园收益。

关键词：玉皇柚采收期授粉果实品质无籽果实有籽果实

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利用InDel标记筛选多胚山金柑珠心苗后代

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果树学报 2023年第7期40卷 1312-1317页

作者：宋谢天田啸宇王楠周银谢源源谢宗周柴利军叶俊丽邓秀新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 湖北洪山实验室武汉430070

【目的】柑橘的遗传转化通常以实生苗上胚轴为外植体。作为基因组高度杂合的无融合生殖物种,柑橘多胚种子中常含有一定比例的、因遗传重组较亲本表现出遗传背景差异的有性后代,从实生群体中筛选无性后代进行遗传转化实验可有效保证外植... 详细信息

【目的】柑橘的遗传转化通常以实生苗上胚轴为外植体。作为基因组高度杂合的无融合生殖物种,柑橘多胚种子中常含有一定比例的、因遗传重组较亲本表现出遗传背景差异的有性后代,从实生群体中筛选无性后代进行遗传转化实验可有效保证外植体材料遗传背景的一致性及后续相关研究的科学性和可靠性。基于此,开发分子标记用于快速准确筛选柑橘短童期种质——多胚山金柑(Fortunella hindsii)子代中的珠心胚实生苗,以期为优化完善山金柑遗传转化体系提供基础。【方法】利用母本山金柑材料DB02重测序数据进行InDel标记的挖掘,筛选出可以区分有性后代和无性后代的InDel标记,通过琼脂糖凝胶电泳对DB02系山金柑的子代幼苗进行鉴定。【结果】开发出7对可以区分DB02系山金柑种子中有性和无性后代的InDel杂合位点标记(InDel1~InDel7),基于这些标记进一步研究,发现土播幼苗和试管幼苗中的珠心苗比例分别为87.5%和74.2%。【结论】利用InDel标记可以成功筛选出与DB02系山金柑遗传背景一致的无性克隆后代,进一步完善了山金柑实生苗上胚轴遗传转化体系。

关键词：山金柑珠心胚实生苗鉴定 InDel 分子标记

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