菊芋果聚糖:果聚糖1-果糖基转移酶1-FFT基因的克隆及表达分析

果聚糖作为一种重要的渗透调节物质,与植物碳素分配和抗逆性的密切相关。为明确果聚糖在植物生长发育过程中的作用,本研究以菊芋为材料,克隆出果聚糖:果聚糖1-果糖基转移酶1-FFT基因,其CDS长2 025 bp,编码641个氨基酸,该蛋白的分子量(Mw)为73.906 k D,蛋白的等电点(p I)为10.01。进化树分析显示菊芋中1-FFT基因与维氏菊的1-FFT基因进化距离最近,具有较高的保守性。荧光定量检测结果显示,1-FFT基因在菊芋的叶、根、花、茎和块茎中均有表达,其中在块茎中表达量最高。本研究为解析果聚糖合成酶的分子调控机理及功能鉴定提供依据。     

甘蓝型油菜PIN家族基因的鉴定与生物信息学分析

PIN家族基因是一类调控植物生长素极性运输的重要载体元件, PIN基因编码生长素输出蛋白, 介导生长素在植物体的运输, 然而在基因组较复杂的甘蓝型油菜中缺乏系统研究。本研究运用生物信息学方法在甘蓝型油菜全基因组数据库筛选甘蓝型油菜PIN家族基因, 对鉴定出的29个BnPINs基因开展拷贝数变异、分子特征、跨膜结构域、保守基序、染色体定位、系统进化树构建、PIN蛋白二级结构及三级结构预测等研究, 结合高通量转录组测序进行低氮胁迫下的转录水平分析。结果表明, 甘蓝型油菜PIN家族基因拷贝数明显多于拟南芥、甘蓝和白菜所具有的PIN家族基因数量; BnPINs蛋白多属于由碱性氨基酸组成的稳定蛋白, 含有保守的N末端结构域, 二级结构与拟南芥PIN蛋白相似; 系统进化选择能力分析表明, BnPINs基因与甘蓝和白菜PIN家族基因进化关系相近。转录组测序表明, BnPIN1s、BnPIN2s、BnPIN3s基因主要在甘蓝型油菜根部表达且受长期低氮(72 h)诱导, BnPIN6s和BnPIN8s基因主要在地上部表达, 低氮会抑制BnPIN6s表达。本研究结果为进一步研究甘蓝型油菜PIN家族基因生物学功能尤其是在响应低氮胁迫中的功能奠定基础, 为已知大量数据的其他物种家族基因生物信息学研究提供参考。     

彩色棉多药和有毒化合物输出蛋白MATE家族基因的鉴定及表达分析

植物多药和有毒化合物输出家族(multidrug and toxic compound extrusion, MATE)是一类可转运阳离子染料、氨基葡糖、多种抗生素与药物等次生代谢产物的转运蛋白家族。本研究利用生物信息学手段从陆地棉基因组数据库中鉴定了MATE家族基因, 并从基因的系统进化关系、染色体分布、基因结构和表达模式等方面对该基因家族特征进行了比较分析。共鉴定出91个陆地棉MATE基因, 命名为GhMATE1~GhMATE91。陆地棉MATE蛋白与拟南芥MATE蛋白均可分为A、B、C、D、E、F和G 7个亚家族, 其中84个GhMATE蛋白具有12个典型的跨膜结构域。染色体定位显示, GhMATE家族成员定位在不同的25条染色体上, 共形成5个基因簇。qRT-PCR分析发现, GhMATE家族基因在棉花各组织中均有表达, 但表达模式各不相同, 其中GhMATE13和GhMATE23在棕色棉纤维中的表达量明显高于在白色棉中, 表明它们可能与棕色棉纤维的颜色形成相关。本研究为进一步解析棉花MATE家族基因的功能和作用机制积累了有价值的资料。     

谷子HAK/KUP/KT钾转运蛋白家族全基因组鉴定及其对低钾和高盐胁迫的响应

KT/HAK/KUP (HAK)家族是植物中最丰富的钾转运体家族,对植物的生长和环境适应具有重要作用。谷子是抗逆耐瘠研究的模式植物,然而,谷子中HAK家族缺乏系统研究。本研究基于基因组序列信息,鉴定出29个谷子HAK基因(SiHAKs),并对该家族成员的基本特征、蛋白结构、染色体定位、基因复制、表达模式和逆境响应等方面进行了系统分析。结果显示,(1) SiHAKs分为5个进化簇(Cluster I～Cluster V),成员数量分别为11、9、3、3和3。基因结构和蛋白保守基序分析表明,谷子HAK家族具有较高的保守性,不同Cluster的保守性依次为:ClusterIII=Cluster V> Cluster II> Cluster I> Cluster IV。(2)串联复制是SiHAKs扩增的主要原因, 15个SiHAKs位于串联重复中。(3)171个转录因子可能结合到不同SiHAKs的启动子上,这些转录因子包含ERF、NAC、MYB和WRKY等家族中的大量成员,可能授予了SiHAKs对非生物胁迫多样的响应机制。(4)基因表达聚类将SiHAKs分成3组:GroupI...     

全基因组鉴定和表达模式分析玉米AP2转录因子

AP2属于AP2/ERF转录因子家族中的一个亚族,其主要特征是具有2个独立的AP2/ERF结构域。本研究利用生物信息学的方法在玉米基因组中鉴定到22个玉米AP2转录因子,并对其理化性质、基因结构、保守基序、系统进化关系和表达模式进行了分析。结果表明,玉米AP2家族成员的理化性质差异较大,且都属于亲水性蛋白。系统进化分析表明玉米AP2家族基因被分为euANT、baselANT、euAP2三个亚组,相同亚组的基因具有相似的基因结构和保守基序。共线性分析发现玉米AP2家族基因中不存在基因串联重复事件,只存在两对片段复制事件,同时结果表明禾本科AP2家族基因之间具有很好的共线性关系。表达模式分析发现ZmAP2基因广泛参与植物的生长发育。转录组数据分析发现,在盐胁迫下一共鉴定到16个ZmAP2基因。本研究为进一步研究玉米AP2转录因子家族的生物学功能提供了科学依据。     

拟南芥NSP家族基因序列分析及响应干旱胁迫表达分析

芥子油苷是十字花科植物中的一种重要的次生代谢产物,它被酶水解的降解过程与植物响应逆境胁迫有关。NSP蛋白是一类能与黑芥子酶结合从而改变芥子油苷降解途径最终生成腈类物质的特异蛋白,其与ESP家族蛋白在功能上存在冗余。本研究以拟南芥NSP家族基因为对象,对家族成员基因序列、蛋白质序列、启动子序列进行生物信息学分析,并使用实时荧光定量PCR技术检测了该家族基因在干旱胁迫下的表达量。结果表明:该家族成员基因分布于3条染色体上,外显子数目2～4个;AtNSP5蛋白亚细胞定位于细胞核,其他成员定位于细胞质;结构域预测显示,该家族蛋白均含有4个Kelch结构域,除AtNSP5外,其他四个成员还含有1～2个Jacalin结构域;启动子序列中均含有CAAT-box、TATA-box核心元件及数量不一的逆境响应元件;干旱胁迫表达量分析显示,该家族中AtNSP5基因受干旱胁迫诱导表达,其他成员响应干旱胁迫不显著。本研究为进一步研究NSP蛋白在植物响应干旱胁迫中的分子机制提供依据,并为植物抗旱基因工程提供潜在候选基因。     

陆地棉MADS-box家族基因鉴定及组织特异性表达分析

【目的】MADS-box蛋白是1种重要的转录因子,参与调控棉花生长发育进程。鉴定分析陆地棉MADS-box家族基因,可为研究其生物学功能奠定基础。【方法】基于2019年最新组装的陆地棉TM-1参考基因组,通过HMMER 3.0软件鉴定陆地棉TypeⅠ型及MIKC型MADS-box基因。利用MapInspect、MEGA7.0、MEME和TBtools软件以及omicshare网站进行染色体定位、多序列比对聚类分析、Motif预测、基因结构鉴定以及组织特异性表达分析。【结果】从全基因组水平上鉴定出181个陆地棉MADS-box基因(66个TypeⅠ型和115个MIKC型),染色体定位发现该181个基因在陆地棉26条染色体上均有分布,其中A组染色体上分布有78个基因、D组上分布有103个基因。系统进化分析显示TypeⅠ型MADS-box蛋白有3个亚家族,MIKC型蛋白包括MIKC*型和含有10个亚家族的MIKCC型;motif预测结果显示所有MADS-box蛋白均含有MADS结构域,基因结构分析结果显示,外显子、内含子结构和长度在各亚家族内具有同一性;组织特异性表达分析发现MADS-box家族基因中有33个在纤维组织中优势表达,103个在花器官中优势表达,41个基因在根茎中优势表达。【结论】本研究通过生物信息学方法,鉴定并获得了与陆地棉开花调控和纤维发育相关的MADS-box基因,对于揭示棉花纤维品质等重要性状的遗传调控机制及分子育种具有一定的理论意义和应用价值。     

玉米TCP转录因子家族的全基因组鉴定及表达模式分析

TCP (Teeosinte branched I, Cycloidea, Proliferating Cell Factors 1和2)是一类植物特有的转录因子家族,包含一个高度保守且非典型的螺旋-环-螺旋(bHLH)结构域,在植物生长发育的不同阶段发挥着重要作用,但在玉米中该基因家族的相关研究报道较少。本研究利用生物信息学方法在玉米全基因组中鉴定TCP转录因子家族成员,并对该家族成员的理化性质、基因结构、染色体定位、进化关系、保守基序及表达模式等进行了分析。结果表明,玉米TCP转录因子家族共包含43个成员,根据系统发育分析可以将其划分为ClassⅠ和ClassⅡ两个亚家族,其中,ClassⅡ亚家族又可以分为CYC-TCP和CIN-TCP两个亚类。玉米TCP转录因子家族蛋白的氨基酸残基数为44～269aa,相对分子质量在5.20～29.54kD,等电点范围在5.04～11.55。基因染色体定位分析发现,玉米10条染色体上均含有TCP基因,且在4号染色体上最多,有8个基因。对该家族基因结构的分析表明,在玉米TCP家族中有66.8%的基因只含有一个外显子,低于拟南芥的82%,表明玉米TCP拥有更加丰富的基因结构。此外,本研究在玉米TCP家族蛋白中共鉴定到20个保守基序,其中有32个TCP蛋白包含motif 1,占总数的74.4%。玉米籽粒RNA-Seq数据分析表明,玉米TCP转录因子家族有26个成员在玉米籽粒中表达,ZmTCP37等基因在不同部位、不同时期均有表达,ZmTCP8等基因在特异部位、特异时期表达。本研究在玉米中鉴定了TCP基因家族基因数量、系统进化关系、保守结构域、在染色体上的定位及玉米籽粒中TCP基因表达模式,为揭示玉米TCP基因家族在玉米籽粒中的功能奠定了基础。     

大白菜NPR家族基因鉴定及表达模式分析

为明确大白菜NPR转录辅助因子在大白菜基因组中的数量、结构和表达特点,揭示NPR在水杨酸信号途径调控大白菜生长发育过程中的作用。本研究利用拟南芥资源信息库TAIR10和芸薹属基因组网站Brassica Database鉴定大白菜基因组中的NPR家族成员,并对其蛋白和基因特性进行生物信息学分析。鉴定出大白菜基因组中包括11个NPR家族基因,分别定位在大白菜7条染色体上,具有1~5个外显子不等。BrNPR家族蛋白可聚类为6个分支,包含11个保守元件,并且所有BrNPR家族成员均含有DUF和Ank两个保守结构域,其中BrNPR1-4成员基因的编码区均含有核定位信号(NLS)。基因表达模式检测结果表明部分大白菜NPR基因在不同组织中具有组织特异的表达模式,BrNPR1B在根、茎和叶及BrNPR3B在花和种荚中表达显著,绝大部分BrNPR基因受盐胁迫诱导表达,部分BrNPR基因受根肿菌侵染诱导表达。本研究可为后续大白菜NPR基因的功能验证提供依据。     

非生物胁迫下白桦ERF基因的表达及生物信息学分析

本研究对3个白桦AP2/ERF家族基因进行了生物信息学分析,并研究了盐和干旱胁迫下白桦ERF基因的表达模式。结果表明,bERF1和bERF2蛋白定位在细胞核内,而bERF3蛋白则定位在叶绿体上;三个基因都含有磷酸化位点;bERF1、bERF2和bERF3基因都属于ERF家族的ERF亚家族,bERF1和bERF2属于B-1组,与拟南芥的AT1G28360.1同源性最高,B-1组的拟南芥ERF基因都属于转录抑制子,说明bERF1和bERF2基因可能具有转录抑制作用;bERF3属于B-5组,与拟南芥的AT4G11140.1同源性最高。3个ERF基因都可以响应盐和干旱胁迫,并且在不同组织中表达量存在差异。     

Potato Tuber Resistance to Phytophthora infestans and its Relation to Maturity

Tuber resistance to P. infestans was evaluated in the progeny of potato genotypes differing in levels of resistance and maturity. Two methods of tuber evaluation were used: testing whole tubers and testing tuber slices. In the progeny, some genotypes were superior in resistance to the more resistant parent. This superiority was never associated with early maturity. The results of both tuber-testing methods were positively correlated but the interaction genotypes X years was more pronounced in the reaction of tuber slices. It is concluded that: (1) transgression in tuber resistance may be obtained, (2) a high level of tuber resistance is difficult to combine with early maturity and (3) the expression of genes for specific resistance was more pronounced in testing tuber slices than in testing whole tubers.     

Effect of Defoliation on Growth and Physiological Developments in Tubers of Purple Nutsedge (Cyperus rotundus L.)

The starch metabolism and hydrolytic enzyme activities in growing tubers of purple nutsedge (Cyperus rotundus L.) in response to defoliation were examined to elucidate the regrowth physiology of the plant. Effects of complete or partial defoliation imposed at 2 or 4 weeks (single defoliation), or 2 and 4 weeks (repeated defoliation) after emergence (WAE) on plant growth were studied.
Leaf area and leaf dry weight per plant in the control plots increased with time. In the mother tuber, dry weight and starch content were reduced by 16 % and 18 %, respectively, during the first 2 WAE, and then increased with time. Endoamylase (EC 3.2.1.1) activity of tubers (3.8 units g⁻¹ dry tuber) increased by 42 % during the first 4 WAE, but decreased rapidly up to 8 WAE.
Repeated defoliation significantly reduced the regrowth ability of nutsedge tubers when compared to single defoliation and the control. Repeated complete defoliation reduced the leaf area and leaf dry weight by 56 % and 69 %, respectively. The dry weights of mother tuber and total tubers 100 cm ⁻² were reduced by 41 % and 63 %, respectively, when compared to the control at 8 WAE. Endoamylase activity and starch content in the mother tuber in this treatment were reduced by 72 % and 56 %, respectively. Defoliation did not have any significant effect on the exoamylase activity.
Endoamylase is the major starch hydrolysing enzyme in tubers and the activity of this enzyme could be a limiting factor in regrowth of nutsedge tubers. Repeated complete defoliation effectively suppressed the regrowth of purple nutsedge.     

春马铃薯种薯大小对其植株性状及产量的影响研究

该文选用过翻秋的小整薯作种进行春播试验，探讨了不同大小种薯对植株性状和产量的影响及其经济效益。结果表明，从植株性状和产量上看，每窝种薯数在１－４范围内，随着种薯数的增加，每窝基本茎数增加，出苗后第３０天冠层覆盖度增加，株高降低，茎粗变细，结薯数增加，埠茎产量变化不大，单个块茎重减少，大中薯数变化不多，大中薯重下降，大中薯数率和大中薯重率降低，其中每窝和薯数与基本茎数，出苗后第３０天的冠覆盖，株高，     

反义PPO基因对马铃薯块茎褐化的影响

对刚收获和贮藏5个月后的12个转基因马铃薯品系及对照块茎进行褐化指标的生理检测,发现2个检测时期的各褐化指标在供试品系间差异均达到显著或极显著水平。综合2个检测时期并与对照相比,PPO活性降幅为26.01%~48.65%;酚物质含量降低22.83%~54.81%;褐化强度除GD-9-qc-1低于对照40.45%~46.78%外,其余品系高于对照或与对照无差别。切割块茎发现,大部分转基因品系较对照褐化出现晚,褐化程度低,褐化指数低于对照88.0%~98.86%。PPO的化学定位显示,低褐化转基因品系的多酚氧化酶集中于维管束环,含量明显少于对照,其中GD-9-qc-1仅有少量多酚氧化酶显现出来,推测是由于反义PPO基因抑制块茎POT32基因表达的结果。进一步的方差、相关分析表明,除对多酚氧化酶含量抑制外,反义PPO基因还降低酚物质含量以减轻块茎损伤褐化。     

叶面喷施MnSO4对马铃薯植株叶片生理特性和块茎品质的影响

锰是马铃薯生长发育不可缺少的微量元素之一,本试验以‘青薯9号’为供试品种,设置了0(CK)、0.05%(T1)、0.1%(T2)、0.6%(T3)、1.1%(T4)5个MnSO₄浓度处理,对马铃薯幼苗期至块茎形成期进行叶面喷施,研究锰素对马铃薯叶片生理特性以及块茎贮藏期品质变化的影响。结果表明,过氧化物酶(peroxidase,POD)、氧化物歧化酶(oxide dismutase,SOD)、过氧化氢酶(catalase,CAT)活性随着喷施MnSO₄浓度的增加呈先升高后降低的趋势,其中喷施浓度为0.05%~0.1%MnSO₄可显著提高马铃薯叶片的POD活性,浓度为0.1%MnSO₄促进SOD、CAT活性的提高。喷施浓度为1.1%MnSO₄有利于块茎形成期根系活力(TTC还原量)的提高。就马铃薯块茎品质而言,喷施浓度为0.1%~0.6%MnSO₄可有效提高马铃薯块茎可溶性蛋白、有机酸、Vc、淀粉含量,且有利于减少贮藏期其各品质的流失;而喷施MnSO₄对块茎有机酸含量影响较小。由此表明,喷施0.1%~0.6%的MnSO₄可提高马铃薯植株叶片生理特性,并优化马铃薯品质。     

Effects of elevated CO2 and/or ozone on nutrient concentrations and nutrient uptake of potatoes

Potato crops were grown at seven sites across Europe to test the effects of elevated atmospheric carbon dioxide and/or tropospheric ozone concentrations on growth, yield and various aspects of potato tuber quality within the framework of the EC funded programme Changing Climate and Potential Impacts on Potato Yield and Quality (CHIP). Field exposure systems were used to enrich the atmosphere in CO₂ and/or ozone. At five of the sites, nutrient element conconcentrations (macronutrients: nitrogen, phosphorus, potassium, calcium, magnesium, and micronutrients: mangenese, zinc, iron) in different parts of plants from the various treatments were analysed. Under elevated CO₂, nearly all nutrient elements tended to decrease in concentration. At maximum leaf area, a significant reduction was observed for the concentrations of nitrogen and potassium both in aboveground biomass and in tubers, and for calcium in tubers. Since CO₂ enrichment promoted early tuber growth, these effects could in part be attributed to tuber developmental stage. At maturity, potato grown under CO₂ enrichment exhibited significantly lower concentrations of nitrogen, manganese and iron in aboveground organs, and of nitrogen, potassium and magnesium in tubers which means a reduction of tuber quality. In contrast to CO₂, elevated ozone tended to increase tuber nutrient element concentrations. This was significant for nitrogen and manganese. CO₂ effects on tuber biomass increase were more pronounced than CO₂ effects on nutrient element decrease. Thus, the total amount of nutrient elements taken up by potato crops increased under elevated CO₂. Fertiliser practice in a future, CO₂-rich world will have to be adjusted accordingly.     

Resistance to Phytophthora infestans in Potato Genotypes Originating from Wild Species

The reaction to P. infestans in the foliage and tubers of potato was evaluated in resistant genotypes and in their progeny. The genotypes originated from wild species known to carry genes for specific resistance. Evidence was obtained that three types of resistance to P. infestans were presented in the genotypes: (1) Resistance due to major, dominant genes was predominant. The presence of these genes was expressed in both foliage and tubers and did not influence maturity. Some of the genes were R genes, providing specific resistance, but for others this could not be determined as they were effective against the fungus inoculum with the highest virulence available: 1.2.3.4.5.7.10.11. (2) Resistance was present, associated with late maturity. (3) Tuber resistance was present, expressed after inoculation of whole tubers but not after inoculation of tuber slices. Testing of the progeny was essential for the determination of the type of resistance present in the genotypes evaluated.     

铵态氮素促进甘薯块根形成的解剖特征及其IbEXP1基因的表达

This study chose Shangshu 19 (S19) and Jixu 23 (J23) categorized by valid tuber root number per plant as analyzing varieties and arranged treatment combinations consisted of two nitrogen forms ammonium nitrogen (A_N) and amide nitrogen (X_N) integrated with two nitrogen rates 60 kg hm^-2 (L_N60) and 180 kg hm^-2 (H_N180), using field and pot trial assays, plus a check treatment received no nitrogen supply in order to make research on the anatomical observation on sweet potato tuber root differentiation and expression characteristics of IbEXP1 gene associated with tuber root formation in ammonia nitrogen in 2014 and 2015. Our results showed that the storage root yield of Shangshu 19 associated with more valid tuber root number per plant was significantly higher than Jixu 23 at harvest stage, which was significant difference compared with each other. In addition, nitrogen levels and nitrogen forms had significant interaction effects. The 60 kg hm^-2 ammonium nitrogen treatment in two sweet potato genotypes achieved the highest final storage root yield in field experiment and showed higher valid tuber root number per plant, which attributed to the younger tubers whose root diameter between 0.5 cm and 5.0 cm during the canopy closure period. It had been observed that 60 kg hm^-2 ammonium nitrogen treatment possessed the most vessels in the primary xylem bundle and the lignified parenchyma cells of the stele tissues in the pre-cambial period, followed with possessing high level expression of IbEXP1 gene, the biggest root diameter and stele diameter and the most number of primary and secondary xylem bundles in the course of primary cambium growth. As the vascular cambium was initiated, relative expression of IbEXP1 gene at 60 kg hm^-2ammonium nitrogen treatment and the degree of parenchyma cells lignification were intermediate between no nitrogen application and high nitrogen treatments, however, the diameter of root and stele and the ratio of them were highest, which achieved the perfect harmony in lignification and division of parenchyma cells in tuberization.     

Differentiation of the high molecular weight glutenin subunit D t x 2.1 of Aegilops tauschii indicated by partial sequences of its encoding gene and SSR markers

The devastating late blight pathogen Phytophthora infestans infects foliage as well as tubers of potato. To date, resistance breeding has often focused on foliage blight resistance, but tuber blight resistance is becoming more and more important in cultivated potatoes. In this study, a reliable tuber assay for resistance assessment was developed and foliage and tuber blight resistance (R) was compared in four mapping populations. In the RH4X-103 population, tuber blight resistance inherited independently from foliage blight resistance. Three specific R genes against P. infestans were segregating. The Rpi-abpt and R3a genes function as foliage-specific R genes, whereas the R1 gene acts on both foliage and tuber. In the segregating populations SHRH and RH94-076, tuber and foliage blight resistance correlated significantly, which suggests that resistance in foliage and tuber is conferred by the same gene (could be R3b) and quantitative trait loci (QTL), respectively. In the CE population neither tuber nor foliage resistance was observed.