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Genome-wide identification of the CONSTANS-LIKE(COL)family and mechanism of fruit senescence regulation by PpCOL8 in sand pear(Pyrus pyrifolia)

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Journal of Integrative Agriculture 2024年第4期23卷 1222-1237页

作者： Yue Xu Shurui Song Huiying Wang Xilong Cao Xinran Zhao Wenli Wang Liyue Huo Yawei Li misganaw wassie Bin Lu Liang Chen Haiyan Shi College of Horticulture Hebei Agricultural UniversityBaoding 071001China Wuhan Botanical Garden Chinese Academy of SciencesWuhan 430074China

Pyrus pyrifolia Nakai‘Whangkeumbae'is a sand pear fruit with excellent nutritional quality and ***,the industrial development of pear fruit is significantly limited by its short shelf *** acid(SA),a well-known phytohormone,can delay fruit senescence and improve shelf ***,the mechanism by which SA regulates CONSTANS-LIKE genes(COLs)during fruit senescence and the role of COL genes in mediating fruit senescence in sand pear are poorly *** this study,22 COL genes were identified in sand pear,including four COLs(Pp COL8,Pp COL9a,Pp COL9b,and Pp COL14)identified via transcriptome analysis and 18 COLs through genome-wide *** COL genes were divided into three subgroups according to the structural domains of the COL *** COL8,with two B-box motifs and one CCT domain,belonged to the first *** contrast,the other three Pp COLs,Pp COL9a,Pp COL9b,and Pp COL14,with similar conserved protein domains and gene structures,were assigned to the third *** four COLs showed different expression patterns in pear tissues and were preferentially expressed at the early stage of fruit ***,the expression of Pp COL8 was inhibited by exogenous SA treatment,while SA up-regulated the expression of Pp COL9a and Pp ***,Pp COL8 interacts with Pp MADS,a MADS-box protein preferentially expressed in fruit,and SA up-regulated its *** the production of ethylene and the content of malondialdehyde(MDA)were increased in Pp COL8-overexpression sand pear fruit,the antioxidant enzyme(POD and SOD)activity and the expression of Pp POD1 and Pp SOD1 in the sand pear fruits were down-regulated,which showed that Pp COL8 promoted sand pear fruit *** contrast,the corresponding changes were the opposite in Pp MADS-overexpression sand pear fruits,suggesting that Pp MADS delayed sand pear fruit *** co-transformation of Pp COL8 and Pp MADS also delayed sand pear fruit *** re

关键词： Pyrus pyrifolia CONSTANS-LIKE gene salicylic acid fruit senescence MADS

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A synthetic light-inducible photorespiratory bypass enhances photosynthesis to improve rice growth and grain yield

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Plant Communications 2023年第6期4卷 85-95页

作者： Huawei Xu Huihui Wang Yanwen Zhang Xiaoyi Yang Shufang Lv Dianyun Hou Changru Mo misganaw wassie Bo Yu Tao Hu College of Agriculture Henan University of Science and TechnologyLuoyang 471000China CAS Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture Wuhan Botanical GardenThe Innovative Academy of Seed DesignChinese Academy of SciencesWuhan 430074China Shanghai Center for Plant Stress Biology CAS Center for Excellence in Molecular Plant SciencesChinese Academy of SciencesShanghai 200032China State Key Laboratory of Herbage Improvement and Grassland Agro-ecosystems College of Pastoral Agriculture Science and TechnologyLanzhou UniversityLanzhou 730020China

Bioengineering of photorespiratory bypasses is an effective strategy for improving plant productivity by modulating *** previouswork,two photorespiratory bypasses,theGOC andGCGT bypasses,increased photosynthetic rates but decreased seed-setting rate in rice(Oryza sativa),probably owing to excess photosynthate accumulation in the *** solve this bottleneck,we successfully developed a newsynthetic photorespiratory bypass(called theGMAbypass)in rice chloroplasts by introducing Oryza sativa glycolate oxidase 1(OsGLO1),Cucurbita maxima malate synthase(CmMS),and Oryza sativa ascorbate peroxidase7(OsAPX7)into the rice genome using a high-efficiency transgene stacking *** the GOC and GCGT bypass genes driven by constitutive promoters,OsGLO1 in GMA plants was driven by a light-inducible Rubisco small subunit promoter(pRbcS);its expression dynamically changed in response to light,producing a more moderate increase in *** rates were significantly increased inGMA plants,and grain yieldswere significantly improved under greenhouse and field *** GMA rice showed no reduction in seed-setting rate under either test condition,unlike previous photorespiratory-bypass rice,probably reflecting proper modulation of the photorespiratory ***,these results imply that appropriate engineering of the GMA bypass can enhance rice growth and grain yield without affecting seed-setting rate.

关键词： photorespiratory bypass photosynthesis rice grain yield seed-setting rates

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Simultaneous gene editing of three homoeoalleles in self-incompatible allohexaploid grasses

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Journal of Integrative Plant Biology 2021年第8期63卷 1410-1415页

作者： Liang Zhang Tao Wang Guangyang Wang Aoyue Bi misganaw wassie Yan Xie Liwen Cao Huawei Xu Jinmin Fu Liang Chen Yang Zhao Tao Hu CAS Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture Wuhan Botanical GardenThe Innovative Academy of Seed DesignThe Chinese Academy of SciencesWuhan430074 China Center of Economic Botany Core Botanical GardensThe Chinese Academy of SciencesWuhan430074 China University of Chinese Academy of Sciences Beijing100049 China Shanghai Center for Plant Stress Biology and CAS Center of Excellence in Molecular Plant SciencesThe Chinese Academy of SciencesShanghai200032 China College of Agriculture Henan University of Science and TechnologyLuoyang471023 China School of Resources and Environmental Engineering Ludong UniversityYantai264011 China

Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system has been widely used for precise gene editing in ***,simultaneous gene editing of multiple homoeoalleles remains challenging,especially in self-incompatible polyploid ***,we simultaneously introduced targeted mutations in all three homoeoalleles of two genes in the self-incompatible allohexaploid tall fescue,using both CRISPR/Cas9 and LbCas12a(LbCpf1)***-of-function mutants of FaPDS exhibited albino leaves,while knockout of FaHSP17.9 resulted in impaired heat resistance in T0 generation of tall ***,these mutations were *** findings demonstrate the feasibility of generating loss-of-function mutants in T0 generation polyploid perennial grasses using CRISPR/Cas systems.

关键词： allohexaploid Cas12a(Cpf1) CRISPR/Cas9 self-incompatible tall fescue

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紫花苜蓿应答高温胁迫及水杨酸提高其耐高温能力的机制研究

紫花苜蓿应答高温胁迫及水杨酸提高其耐高温能力的机制研究

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作者： misganaw wassie Anley 中国科学院大学(中国科学院武汉植物园)

学位级别：硕士

紫花苜蓿（Medicago sativa L.）是世界上栽培和应用最广泛的牧草,具有营养价值丰富、产量高等特点,素有“牧草之王”的美誉,主要集中分布在我国西北、东北、华北等地。随着我国南方草食畜牧业的发展,对高品质的青绿饲料需求也越来越大... 详细信息

紫花苜蓿（Medicago sativa L.）是世界上栽培和应用最广泛的牧草,具有营养价值丰富、产量高等特点,素有“牧草之王”的美誉,主要集中分布在我国西北、东北、华北等地。随着我国南方草食畜牧业的发展,对高品质的青绿饲料需求也越来越大,紫花苜蓿作为优质的豆科牧草也逐渐从北向南推广种植。紫花苜蓿喜温凉,在生长季节内易受到南方夏季高温影响,导致产量和品质下降。因此,研究紫花苜蓿响应高温胁迫的生理分子机制具有重要科学意义。在此,我们首先研究了高温胁迫对不同品种紫花苜蓿表型及生理指标的影响,并筛选出了紫花苜蓿耐高温品种及高温敏感品种;随后,以耐高温紫花苜蓿品种为材料,通过转录组分析探究了紫花苜蓿响应高温胁迫的分子机制;最后,通过外施水杨酸（SA）探究了外源水杨酸在缓解紫花苜蓿高温胁迫损伤中的作用。本论文取得的研究结果如下。1.高温胁迫对紫花苜蓿生长和生理特性的影响为了探究高温胁迫对紫花苜蓿生理指标的影响,对15个品种的4周苗龄紫花苜蓿进行38/35℃高温处理,并测定高温处理7天后幼苗的各项生长和生理指标。结果表明,高温胁迫显著降低了大部分紫花苜蓿品种的生物量、相对含水量（RWC）和叶绿素含量,增加了电解质渗漏率（EL）、丙二醛含量（MDA）、可溶性糖含量（SS）和可溶性蛋白（SP）含量。根据隶属函数值,‘Bara310SC’（0.86）和‘Magna995’（0.80）为耐高温品种,‘Gibraltar’（0.24）和‘WL712’（0.24）为高温敏感品种。进一步地,对上述筛选出的4个品种紫花苜蓿进行叶绿素荧光分析,发现高温对Bara310SC’的光合活性影响最小,反之,对WL712’的光合活性影响最大。上述结果表明,‘Bara310SC’和‘WL712’分别为紫花苜蓿耐高温品种和高温敏感品种。2.高温胁迫下紫花苜蓿的转录组分析为了探究紫花苜蓿响应高温胁迫的分子机制,以紫花苜蓿耐高温品种Bara310SC为材料,对其4周苗龄的幼苗进行38℃高温处理4 h小时,并通过RNA-seq对高温处理前后的叶片进行转录组测序。结果显示,6个紫花苜蓿cDNA文库一共获得177278个unigenes。与对照相比,高温处理的叶片中有4649个基因的表达量发生了显著的变化。其中,2406个基因的表达量受高温显著诱导,而2243个基因的表达量受高温显著抑制。通过对随机选取的10个差异表达基因进行qRT-PCR分析,验证了RNA-seq结果的正确性。随后,对响应高温胁迫的差异基因进行GO（Gene Ontology）和KEGG（Kyoto Encyclopedia of Genes and Genomes）分析。GO分析表明这些差异基因主要富集在‘Protein metabolic process’,‘Oxidoreductase activity’‘Cytosolic part’。KEGG分析发现‘ribosome’,‘oxidative phosphorylation’,‘aminoacyltRNA biosynthesis’和‘citrate（TCA）cycle’是响应高温胁迫的主要通路。3.高温胁迫下SA对紫花苜蓿的保护作用为了探究外源SA在缓解高温对紫花苜蓿的伤害中的作用,我们对4周苗龄的高温敏感紫花苜蓿‘WL712’喷施0.25 mM或0.5 mM的SA,再对SA预处理5 d的紫花苜蓿进行高温胁迫（38/35°C）处理,并测定高温处理后幼苗的各项生长和生理指标。结果表明,在高温胁迫条件下,与对照相比,外源SA预处理可以改善紫花苜蓿的叶片形态,提高株高、生物量、叶绿素含量和光合效率,降低电解质渗透率（EL）和丙二醛（MDA）含量,从而减轻高温诱导的膜损伤。此外,外源SA预处理还能调节过氧化氢酶（CAT）、超氧化物歧化酶（SOD）和过氧化物酶（POD）等抗氧化酶活性。总而言之,外源SA提高了紫花苜蓿的耐高温能力,且较低浓度（0.25 mM）的SA效果更为显著。综上所述,本论文的研究为了解紫花苜蓿响应高温胁迫的生理分子机制提供了基础信息。此外,本研究还为揭示SA介导的紫花苜蓿对高温胁迫的生理适应机制以及外源SA可用于缓解高温诱导对紫花苜蓿的伤害程度提供了基础认识,下一步研究将探索响应候选基因或转录因子在提高紫花苜蓿耐性方面的潜在作用。

关键词：紫花苜蓿热胁迫生理学水杨酸转录组图谱

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