DIA-based proteome profiling with PRM verification reveals the involvement of ER-associated protein processing in pollen abortion in Ogura CMS cabbage

Ogura cytoplasmic male sterility(Ogura CMS)is extensively applied in hybrid seed production in cruciferous crops.However,the posttranscriptional molecular basis of Ogura CMS in cruciferous crops remains elusive.Here,a data-independent acquisition-based proteomic approach coupled with a parallel reaction monitoring-based targeted proteomic assay was used to analyze the proteome dynamics of Ogura CMS cabbage line RM and its maintainer line RF during floral bud development to obtain insights into the mechanism underlying Ogura CMS in cruciferous crops.A total of 9162 proteins corresponding to 61464 peptides were identified in RM and RF floral buds.The proteomic fluctuation of RM was weaker than that of RF.Differences in protein expression between RM and RF gradually enlarged with floral bud development.Fifteen continually up-regulated and eight continually down-regulated proteins were found in RM relative to RF throughout floral bud development.Differentially expressed proteins between RM and RF during floral bud development were implicated in the endoplasmic reticulum(ER)-associated protein processing pathway,in which most of them exhibited down-regulated expression in RM.These data suggest that ER-associated protein processing may be involved in pollen abortion in Ogura CMS cabbage by inhibiting the expression of critical factors.Our findings not only deepen the understanding of the molecular mechanisms of Ogura CMS in cruciferous crops but also provide better guidance for applying Ogura CMS in the hybrid breeding of cruciferous crops.

Advances in DNA methylation and its role in cytoplasmic male sterility in higher plants

The impact of epigenetic modifications like DNA methylation on plant phenotypes has expanded the possibilities for crop development.DNA methylation plays a part in the regulation of both the chromatin structure and gene expression,and the enzyme involved,DNA methyltransferase,executes the methylation process within the plant genome.By regulating crucial biological pathways,epigenetic changes actively contribute to the creation of the phenotype.Therefore,epigenome editing may assist in overcoming some of the drawbacks of genome editing,which can have minor off-target consequences and merely facilitate the loss of a gene’s function.These drawbacks include gene knockout,which can have such off-target effects.This review provides examples of several molecular characteristics of DNA methylation,as well as some plant physiological processes that are impacted by these epigenetic changes in the plants.We also discuss how DNA alterations might be used to improve crops and meet the demands of sustainable and environmentally-friendly farming.

Fine mapping and cloning of the sterility gene Bra2Ms in nonheading Chinese cabbage(Brassica rapa ssp.chinensis)

The application of a male-sterile line is an ideal approach for hybrid seed production in non-heading Chinese cabbage(Brassica rapa ssp.chinensis).However,the molecular mechanisms underlying male sterility in B.rapa are still largely unclear.We previously obtained the natural male sterile line WS24-3 of non-heading Chinese cabbage and located the male sterile locus,Bra2Ms,on the A2 chromosome.Cytological observations revealed that the male sterility of WS24-3 resulted from disruption of the meiosis process during pollen formation.Fine mapping of Bra2Ms delimited the locus within a physical distance of about 129 kb on the A2 chromosome of B.rapa.The Bra039753 gene encodes a plant homeodomain(PHD)-finger protein and is considered a potential candidate gene for Bra2Ms.Bra039753 was significantly downregulated in sterile line WS24-3 compared to the fertile line at the meiotic anther stage.Sequence analysis of Bra039753 identified a 369 bp fragment insertion in the first exon in male sterile plants,which led to an amino acid insertion in the Bra039753 protein.In addition,the 369 bp fragment insertion was found to cosegregate with the male sterility trait.This study identified a novel locus related to male sterility in non-heading Chinese cabbage,and the molecular marker obtained in this study will be beneficial for the marker-assisted selection of excellent sterile lines in non-heading Chinese cabbage and other Brassica crops.

Morphological and cytological assessments reveal pollen degradation causes pollen abortion in cotton cytoplasmic male sterility lines

Background Understanding the mechanism of male sterility is crucial for producing hybrid seeds and developing sterile germplasm resources.However,only a few cytoplasmic male sterility(CMS)lines of cotton have been produced due to several challenges,like inadequate variation of agronomic traits,incomplete sterility,weak resilience of restorer lines,and difficulty in combining strong dominance.Therefore,the morphological and cytological identification of CMS in cotton will facilitate hybrid breeding.Results Two F_(2) segregating populations of cotton were constructed from cytoplasmic male sterile lines(HaA and 01A,maternal)and restorer lines(HaR and 26R,paternal).Genetic analysis of these populations revealed a segregation ratio of 3:1 for fertile to sterile plants.Phenotypic analysis indicated no significant differences in traits of flower bud development between sterile and fertile plants.However,sterile plants exhibited smaller floral organs,shortened filament lengths,and anther atrophy on the flowering day in comparison with the fertile plants.When performed scanning electron microscopy(SEM),the two F_(2) populations revealed morphological variations in the anther epidermis.Cellular analysis showed no significant differences in pollen development before pollen maturation.Interestingly,between the pollen maturation and flowering stages,the tapetum layer of sterile plants degenerated prematurely,resulting in abnormal pollen grains and gradual pollen degradation.Conclusion The results of this study suggest that fertility-restoring genes are controlled by a single dominant gene.Sterile plants exhibit distinctive floral morphology,which is characterized by stamen atrophy and abnormal anthers.Pollen abortion occurs between pollen maturity and flowering,indicating that premature tapetum degradation may be the primary cause of pollen abortion.Overall,our study provides a theoretical basis for utilizing CMS in hybrid breeding and in-depth investigation of the dominant configuration of cotton hybrid combinations,mechanisms of sterility,and the role of sterile and restorer genes.

大白菜Ogura胞质雄性不育恢复材料的创制与检测

针对白菜类蔬菜(Brassica rapa L.)中缺乏Ogura胞质雄性不育育性恢复材料的问题,利用远缘杂交技术将恢复基因Rfo从甘蓝型油菜导入大白菜中,利用种间杂交种不断与不育系进行回交,已分别获得BC_(1)、BC_(2)和BC_(3)代育性恢复材料。同时,利用形态学、细胞遗传学、分子生物学手段对所创制的材料进行检测,利用恢复基因Rfo特异性引物扩增,最后筛选获得了一株育性稳定、形态特征接近大白菜、染色体数目为20的BC_(3)单株。利用所创制BC_(3)单株进行Ogura雄性不育大白菜商品杂交种恢复测试,结合表型及恢复基因Rfo、不育基因orf138特异性引物检测,结果表明,所创制恢复系恢复基因有效。

通过遗传转化Rfo获得青花菜Ogura CMS恢复系

【目的】将来自萝卜的胞质不育恢复基因Rfo导入到青花菜Ogura细胞质雄性不育(cytoplasmic male sterility,CMS)主栽商业品种‘耐寒优秀’(命名为SFB45)中,获得Ogura CMS恢复系,打破Ogura CMS材料在生产上无法再利用的现状,为优异种质的创制和改良提供新途径。【方法】利用基因合成的方法克隆萝卜Rfo的CDS及其启动子序列,构建表达载体pRfo::Rfo。通过农杆菌介导的遗传转化方法侵染SFB45的具柄子叶和下胚轴,进行再生、抗性苗筛选和PCR鉴定获得阳性植株,开花期观察转基因植株的育性恢复情况;用亚历山大染液分析对照组及阳性株的花粉活力;分别提取对照组及阳性株<3 mm和>3 mm花蕾的RNA并反转录成单链cDNA,设计特异引物,利用RT-PCR分析Rfo及绒毡层和花粉壁发育相关基因在SFB45及对应的育性恢复材料花蕾中的表达差异。【结果】通过遗传转化共获得10个阳性株系,其中8个株系育性得到不同程度的恢复,花粉活力为84.2%—90.4%。RT-PCR分析结果表明,育性恢复植株花蕾中Rfo表达,绒毡层发育关键调节因子DYT1和TDF1及花粉壁主要成分孢粉素合成所必需的基因ACOS5在育性恢复植株<3 mm花蕾中上调表达。绒毡层降解相关基因AMS、四分体胼胝质壁及花粉外壁发育所必需的基因CalS5和CYP703在育性恢复植株>3 mm花蕾中上调表达。分析阳性株系R-1、R-3和R-6自交及杂交后代发现转入的Rfo能稳定遗传,符合孟德尔遗传。【结论】通过遗传转化Rfo获得了优良青花菜Ogura CMS商品种SFB45的育性恢复系,且Rfo的转入使绒毡层及花粉壁发育相关基因恢复正常表达。

萝卜胞质雄性不育(Ogura CMS)芥菜材料的创制

为获得萝卜胞质雄性不育芥菜,利用远缘杂交将芥菜的B基因组导入白菜,通过连续回交法结合胚挽救技术,创制出保留白菜雄性不育基因的芥菜。用萝卜胞质雄性不育白菜CCR17001与可育叶用芥菜JC17000杂交,并以叶用芥菜为轮回亲本连续进行5代回交,获得萝卜胞质雄性不育芥菜材料,对其进行形态学、细胞学、分子生物学鉴定。结果表明:该材料植株形态与轮回亲本相似,蜜腺发育良好,雄蕊高度退化成花瓣,不育度和不育株率均为100%,且不育性稳定,有正常结籽能力,可作为芥菜优势育种的可靠材料。

Genic male and female sterility in vegetable crops

Vegetable crops are greatly appreciated for their beneficial nutritional and health components.Hybrid seeds are widely used in vegetable crops for advantages such as high yield and improved resistance,which require the participation of male(stamen)and female(pistil)reproductive organs.Male-or female-sterile plants are commonly used for production of hybrid seeds or seedless fruits in vegetables.In this review we will focus on the types of genic male sterility and factors affecting female fertility,summarize typical gene function and research progress related to reproductive organ identity and sporophyte and gametophyte development in vegetable crops[mainly tomato(Solanum lycopersicum)and cucumber(Cucumis sativus)],and discuss the research trends and application perspectives of the sterile trait in vegetable breeding and hybrid production,in order to provide a reference for fertility-related germplasm innovation.

Jasmonic acid-mediated stress responses share the molecular mechanism underlying male sterility induced by deficiency of ZmMs33 in maize

Plant male reproduction is a fine-tuned developmental process that is susceptible to stressful environments and influences crop grain yields.Phytohormone signaling functions in control of plant normal growth and development as well as in response to external stresses,but the interaction or crosstalk among phytohormone signaling,stress response,and male reproduction in plants remains poorly understood.Cross-species comparison among 514 stress-response transcriptomic libraries revealed that ms33-6038,a genic male sterile mutant deficient in the Zm Ms33/Zm GPAT6 gene,displayed an excessive drought stress-like transcriptional reprogramming in anthers triggered mainly by disturbed jasmonic acid(JA)homeostasis.An increased level of JA appeared in Zm Ms33-deficient anthers at both meiotic and postmeiotic stages and activated genes involved in JA biosynthesis and signaling as well as genes functioning in JA-mediated drought response.Excessive accumulation of JA elevated expression level of a gene encoding a WRKY transcription factor that activated the Zm Ms33 promoter.These findings reveal a feedback loop of Zm Ms33-JA-WRKY-Zm Ms33 in controlling male sterility and JA-mediated stress response in maize,shedding light on the crosstalk of stress response and male sterility mediated by phytohormone homeostasis and signaling.

低温等离子体技术在食品行业的应用研究进展

近年来,低温等离子体技术发展迅速,由于低温等离子体设备具有易安装、样品处理简单、绿色环保等性能优势,因此在食品工业领域越来越受到重视。该文在阐析低温等离子体技术的相关概念及分类的基础上,重点对低温等离子体在食品杀菌与贮存、降解真菌毒素与农药残留及菌藻诱变方面的应用进行阐述。以期为低温等离子体技术在食品行业的广泛应用提供参考。

信息化质量追溯系统在消毒供应中心医疗器械管理中的应用效果

目的:探讨信息化质量追溯系统在消毒供应中心(CSSD)医疗器械管理中的应用效果。方法:选取郑州儿童医院CSSD采取常规管理期间(2020年6月至2021年6月)接收的1018件器械为对照组,选取应用信息化质量追溯系统管理后(2021年7月至2022年7月)接收的1018件器械为观察组。比较两组医疗器械管理质量(清洗、灭菌、包装不合格率)及各科室医护人员对CSSD的满意度。结果:观察组医疗器械清洗、包装、灭菌不合格率均低于对照组,差异有统计学意义(P<0.05);各科室医护人员对观察组服务态度、技术水平、器械供应及时性、器械结构功能、器械无菌性等方面的满意度均高于对照组,差异有统计学意义(P<0.05)。结论:信息化质量追溯系统用于CSSD医疗器械管理可提高管理质量,降低医疗器械清洗、包装、灭菌不合格率,提高各科室医护人员满意度,有效保障医疗安全。

水稻不育系泰丰A创制及其优良品质性状遗传基础研究

针对杂交稻“高产难优质”的瓶颈问题,基于杂种优势群与优势模式,选择华南地区高配合力、高异交率保持系博B和优质丝苗米亲本米31以及长江流域优质抗病早稻浙9248作为育种亲本进行复合杂交,通过穿梭种植生态加压表型鉴定与室内品质鉴定相结合的方法,育成了粒型细长、整精米率高、食味佳的“三高一优”不育系及其保持系泰丰A/B。截至2023年,用泰丰A配组育成泰丰优208、泰优390、泰优1002和泰优398等优质杂交稻共计92个。其中,80.4%的米质达到国(部)标优质3级以上标准,18个品次获得国家或省级优质稻食味鉴评金奖和银奖,2个品种被农业农村部认定为超级稻,较好解决了杂交稻“高产难优质”的“卡脖子”问题。此外,对控制泰丰A/B重要品质性状的QTLs(基因)进行了分析,发现位于第3、5、6和7号染色体上的gs3、GW5TFB、WxTFB和GW7TFB基因或其所在的染色体区段是泰丰A/B品质表现优异的重要遗传基础。

大豆细胞质雄性不育遗传基础与育种应用

杂种优势利用是显著提高作物产量的主要途径,有助于解决日益增长的人口数量与有限耕地之间的矛盾。大豆作为世界上重要的粮油饲兼用作物,其开展杂种优势利用已有30余年。其中,基于细胞质雄性不育的三系法杂交育种系统是大豆杂种优势利用的主要途径。目前,已有40余个杂交大豆品种通过审定并在生产上推广应用,杂交大豆正处于由中试向产业化推进阶段。本文对大豆细胞质雄性不育遗传基础与育种应用进行了综述,系统阐述了各类型细胞质雄性不育系的发现及利用、不育性状的遗传和分子机制、育性恢复基因和恢复抑制基因的定位和克隆等方面的研究进展。基于大豆杂种优势利用研究现状论述和分析,提出了三系法杂交大豆育种中存在的问题、挑战及解决路径,并对三系法杂交大豆育种技术的创新进行了展望,旨在为大豆杂种优势分子基础和应用研究提供新方法、新思路。

基于MPGW分布的压力蒸汽灭菌器可靠性分析

浴缸形状的失效率函数因其能够捕捉某些实际情境中的故障率特征,在机械结构可靠性分析中至关重要。引入一种修正广义幂威布尔(Modified Power Generalized Weibull,MPGW)分布,能够建模表征浴缸形状的失效率变化规律。为评估该模型的有效性,本研究使用了压力蒸汽灭菌器故障数据集进行实证研究,并将其性能与5种经典分布模型进行了比较,发现该模型在灵活性上具有较大优势。

电极间隙对脉冲电场处理器杀菌效果的影响

脉冲电场是新型非热杀菌技术,杀菌效率高、有效保留液体食品的营养成分,具有广阔的应用前景。电极间隙作为脉冲电场杀菌处理器的关键指标,决定处理器的电场分布和杀菌效果。为了获得最佳间隙参数,提升杀菌效果,为此设计了共场型脉冲电场处理器,电极间距分别为3、5和7 mm,针对固定电导率(等效鲜榨柚子汁电导率)的2种典型细菌(金黄色葡萄球菌和大肠杆菌)菌悬液,研究了脉冲电场的杀菌效果。研究发现,3 mm电极间距所能施加最大电压为18 kV,5、7 mm的可达30 kV;处理时间为120 s时,3种间隙的杀菌效率均可达99.99%。因此,综合考虑电源性能、设备能耗和处理能力,首选3 mm电极间距为处理器的应用参数。评估外施电压幅值对杀菌效果的影响,发现细菌存活量的下降趋势可根据其下降速率分为“慢-快-慢”3个阶段。结合菌悬液上清液蛋白质含量的上升趋势,充分说明了电穿孔应为脉冲电场的主要杀菌机制。

西瓜雄性不育类型的遗传学鉴定与胚胎学观察

为了明确西瓜雄性不育材料的不育类型和花药败育时期,以西瓜雄性不育系GMS4为材料,对花蕾不育性状进行田间观察统计,利用石蜡切片法观察西瓜花药发育过程。结果表明,该材料不育性状由1对隐性基因控制,为细胞核雄性不育类型。可育株雄蕊发育正常,不育株雄蕊较小,花药干瘪无花粉,可育株和不育株雌蕊发育无明显差异,均可正常结瓜。从花药发育的整个过程来看,在花药发育早期,可育株和不育株花药结构形态差别不大,在小孢子母细胞减数分裂期,不育株异常绒毡层细胞过度增殖,体积小,排列不整齐,挤压药室,无二分体和四分体形成,在花药发育后期,无花粉粒形成,最终导致西瓜雄性不育发生。据此认为,花药败育发生时期在小孢子母细胞减数分裂期。

茄子反向温敏雄性不育系内源激素含量及相关基因表达分析

雄性不育对杂交育种至关重要,挖掘茄子雄性不育关键候选基因有助于定向改良不育材料。以长茄温敏型雄性不育系05ms及其同源可育系S63为材料,对不育系05ms和可育系S63减数分裂时期(Ⅱ)的花药进行转录组测序,并对05ms春季低温不育(ML)和夏季高温可育(MH)阶段的花粉母细胞时期(Ⅰ)、减数分裂期(Ⅱ)、小孢子时期(Ⅲ)和成熟花粉粒(Ⅳ)4个花药发育时期的内源激素含量进行检测。结果表明,差异表达基因主要富集在戊糖和葡萄糖酸酯的转化、类胡萝卜素的生物合成和植物激素信号转导等代谢通路,Smechr0702283、Smechr0801802、Smechr0600105、Smechr0802213、Smechr0302844、Smechr0601316、Smechr0401804等基因为茄子温敏雄性不育的关键调控基因,实时荧光定量PCR(qRT-PCR)结果表明,转录组数据可靠。激素代谢物检测结果表明,生长素类激素ICAld和乙烯前体ACC含量在4个时期中ML均显著高于MH,而ML的生长素IAA含量低于MH。从代谢水平和转录水平解析了茄子花药的败育机制,研究结果为进一步揭示茄子温敏雄性不育调控机制奠定了理论基础。

辣椒雄性不育分子研究进展

为对辣椒雄性不育分子研究取得的进展有更直观、系统、深入的了解,梳理已鉴定的辣椒核雄性不育(NMS)基因及部分NMS基因分子标记开发,重点阐述近年来核雄性不育基因精细定位及候选基因探究工作;同时回顾辣椒胞质雄性不育(CMS)相关的线粒体不育基因及其关联分子标记和CMS恢复基因分子标记开发,重点概述近年来恢复基因精细定位研究取得的进展。研究指出,精细定位NMS基因,明确候选基因并开发功能标记是辣椒NMS研究的重点,既能为探明辣椒NMS机理奠定基础又可为育种提供高效利用的分子标记。而CMS相较于NMS更为复杂,表现在遗传机制多样且更易受环境影响等方面,因此,从对CMS育性恢复基因的精细定位到基因克隆以及功能研究并探明CMS分子机制还有很长的一段路要走。

适合机械化移栽结球甘蓝新品种‘沪甘2150’的选育

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