Cd2+对番茄幼苗生长和蛋白质组的影响

以3 d龄番茄幼苗为试验材料, 从生理、生化和蛋白质组角度, 分析0.01~1.00 mmol L^–1 Cd²⁺处理72 h后对幼苗的影响。结果表明, Cd²⁺处理导致幼苗生长严重受抑, 幼苗高度从对照组的4.76±0.50 cm分别降至3.79±0.05 cm (0.01 mmol L^–1 Cd²⁺处理, P<0.01)和1.77±0.15 cm (0.03 mmol L^–1 Cd²⁺处理, P<0.01)。根长度从对照组的6.07±0.04 cm降至4.77±0.58 cm (0.01 mmol L^–1 Cd²⁺处理, P< 0.01)和3.65±0.66 cm (0.03 mmol L^–1 Cd²⁺处理, P<0.01)。叶绿素含量在0.1 mmol L^–1 Cd²⁺处理后开始下降。当幼苗用0.05 mmol L^–1 Cd²⁺处理时, 根系中有10个蛋白质斑点, 叶片中有21个蛋白质斑点产生变化。利用MS/MS技术, 根系中有4个蛋白质斑点得以鉴别, 它们是ribosomal protein L 20 (斑点1)、F-box /LRR repeat protein (斑点2)、ribosomal protein small submit 4 (斑点4)和CBL-interacting protein kinase (斑点5)。在叶片中, 有2个蛋白质斑点消失, 4个蛋白质斑点合成, 它们是ABC transporter (斑点16)、maturase-like protein (斑点17)、chalcone synthase (斑点1)、a hypothetical protein (斑点3)、an unknown protein (斑点4)和a predicated protein (斑点6)。这些被鉴别的Cd²⁺反应蛋白参与生物合成、mRNA转录调控和蛋白质转运。     

栽培小麦Brock和京411感染白粉菌后蛋白质组的变化

用华北地区流行的白粉菌15号生理小种,感染强抗白粉病的栽培小麦Brock和对白粉病敏感的小麦京411,通过蛋白质组技术分析其差异蛋白。结果表明,Brock经白粉菌感染12 h后,至少有6个蛋白质斑点(43 kD/pI 6.7、43 kD/pI 6.9、43 kD/pI 7.2、28 kD/pI 5.8、26 kD/pI 5.5和26 kD/pI 6.5)表达量明显增加;感染3 d后,有5个蛋白质斑点(48 kD/pI 5.6、43 kD/pI 6.9、43 kD/pI7.2、28 kD/pI5.8和26 kD/pI5.5)表达量增加;感染5 d后,有12个新的蛋白质斑点(16 kD/pI 7.6、42 kD/pI 6.5、40 kD/pI 4.8、40 kD/pI 4.6、31 kD/pI 5.7、16 kD/pI 4.6、20 kD/pI 8.3、50 kD/pI 6.7、48 kD/pI 6.6、28 kD/pI 5.7、23 kD/pI 4.8和25 kD/pI 4.7 )被诱导合成,2种蛋白质斑点(26 kD/pI 4.6和17 kD/pI 7.9)消失。京411经白粉菌感染12 h后,3个蛋白质斑点(21 kD/pI 6.4、18 kD/pI 5.4和14 kD/pI 7.0)表达量增加;感染3 d后,有2个蛋白质斑点(80 kD/pI 5.4和14 kD/pI 7.0)表达量增加,1个蛋白质斑点(16 kD/pI 5.4)表达量下降;感染5 d后,有3个蛋白质斑点(50 kD/pI 7.3、40 kD/pI 7.3和24 kD/pI 7.2)表达量增加,2个斑点(40 kD/pI 4.8和14 kD/pI 7.2)表达量下降,但没有发现新的蛋白质合成。对Brock中诱导产生的12个新蛋白质斑点,利用MALDI-TOF-MS方法,于NCBI进行数据查询,其中有6个分别属于F-box亮氨酸高度重复蛋白、重金属转运/解毒蛋白、β-1,3-葡聚糖酶(两个同工体)、β-1,3-葡聚糖酶前体、锌指蛋白。功能查询表明,上述6个蛋白参与细胞周期调控、发育、激素响应、基因转录和病害防御等。推测Brock和京411感染白粉菌后,出现的蛋白质组变化可能与各自的抗、感白粉病特性有关。     

赤霉素对盐胁迫抑制水稻种子萌发的缓解作用的蛋白质组分析

用粳稻日本晴(Oryza sativa L. cv.Nipponbare),研究了盐胁迫对水稻种子萌发的抑制作用和赤霉酸(GA₃)对盐胁迫的缓解作用;分别以H₂O (对照）,5 g L^-1 NaCl (处理I),5 g L^-1 NaCl + 100 μmol L^-1 GA₃(处理II)培养水稻种苗48 h,提取芽中的蛋白质,利用双向电泳(2-DE)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术分析了水稻蛋白质组的变化。结果表明,在盐胁迫条件下,日本晴种子的萌发显著受到抑制,而GA₃能显著缓解这种抑制作用;用ImageMaster软件分析2-DE凝胶,发现有4个蛋白质斑点表现出显著的变化,在盐胁迫下斑点S1、S2和S3表达下调而斑点S4消失,在GA₃与盐共处理时,这4个蛋白质点的表达均有不同程度的恢复;经MALDI-TOF MS分析,其中2个蛋白质斑点(S1,S3)分别被鉴定为isoflavone reductase-like蛋白与葡萄糖磷酸变位酶,这些蛋白可能与GA₃提高水稻耐盐性途径相关。     

小麦蛋白质含量遗传规律及品质改良途径研究

本文研究了小麦籽粒蛋白质含量在F1及F2表现中的表现情况。结果表明，控制蛋白质含量的基因作用以加性效应为主。同时存在非加性效应：F1代子粒蛋白质含量与双亲平均值高度相关，F2代中子粒蛋白质含量分离呈正态分布，且正态分布的峰值蛋白质含量也接近双亲平均值。蛋白质含量的一般配合力大于特殊配合力方差，蛋白质含量的广义和狭义遗传分别为79.88%，和49.07%。     

亚麻木质素合成关键酶基因表达分析

以亚麻(Linum usitatissimum)栽培品种白花为材料,采用半定量RT-PCR方法对亚麻木质素合成关键酶基因CCoAOMT1、4CL1、4CL2、COMT、F5H1、F5H2和F5H3在不同组织及不同时期木质部、韧皮部的表达规律进行了研究。结果显示, 除F5H1在根部未表达外,其他基因在幼苗、根、花期木质部、花期韧皮部、叶、花和幼果中均有不同程度表达。CCoAOMT1在各部位都有较高的表达。2个4CL之间和3个F5H基因之间的部位表达的差异存在一定互补性。在韧皮部中,各基因均在幼嫩(30 d)阶段表达量最低;除4CL2在花期(60 d)达到表达高峰外,其他基因均在花后(70 d)表达量最高;4CL1在各时期均有较强表达,而F5H3在各时期弱表达。在木质部中,除4CL1在各时期均表达弱和F5H1在花前期为表达高峰外,其余基因均在幼嫩阶段表达量最高;随后逐渐降低,在花期后表达量大幅降低;与其他基因的明显区别在于各时期4CL1的表达量以韧皮部明显高于木质部。     

铜诱导下小麦生理生化及叶绿体蛋白质组分变化

采用CuCl_2处理小麦幼苗的方法,研究重金属元素铜对小麦幼苗生理生化及蛋白组分影响.结果表明:随着环境中Cu~(2+)浓度的增加小麦苗株高呈显著降低趋势,各浓度处理的苗高均极显著低于对照,5 mmol/L处理较对照降低了54.47%;叶绿素含量呈现降-升-降的趋势,但均小于对照;可溶性蛋白含量分别比对照降低了0.68～1.23 mg/g,但差异不显著.利用双向电泳技术分析了1 mmol/L Cu~(2+)处理后叶绿体蛋白质组分变化,结果表明,有17个蛋白质斑点发生较明显变化,其中7个斑点缺失,1个斑点增加,另有9个蛋白质斑点表达量降低.     

小麦蛋白质含量遗传规律及品质改良途径研究

研究了小麦籽粒蛋白质含量在F1及F2代中的表现情况。结果表明,控制蛋白质含量的基因作用以加性效应为主,同时存在非加性效应;F1代子粒蛋白质含量与双亲平均值高度相关,F2代中子粒蛋白质含量分离呈正态分布,且正态分布的峰值蛋白质含量也接近双亲平均值。蛋白质含量的一般配合力大于特殊配合力方差,在优质育种的亲本选配上,一般配合力更重要一些,应重视蛋白质高配合力亲本的鉴定应用,并计算了蛋白质含量的广义和狭义遗传力。同时对蛋白质数量改良途径进行了分析。     

强筋小麦新麦18号系谱和品质性状遗传分析

对新麦18号的品质性状、农艺性状及其亲缘关系进行了遗传分析。结果表明。新麦18号品质性状属双亲互补且超亲遗传，年际间品质性状稳定。在产量与品质两个负相关性状选育上有重大突破。新麦18号遗传基础丰富．农艺性状继承了双亲的优良性状，5＋10优质亚基和含有偏凸山羊草的抗病基因来源于母本（C5/3577）F3d。     

玉米种子休眠相关蛋白的鉴定与分析

以强休眠性玉米自交系08-641为材料, 2个弱休眠性玉米自交系为对照, 利用蛋白质双向电泳技术, 对处于休眠状态下的新鲜收获种子和经过15 d后熟处理破除休眠的3个自交系种子进行了蛋白质差异表达分析。结果表明, 在3次重复试验下共检测到9个与休眠相关的蛋白点, 其中新增诱导表达蛋白质1个, 缺失表达蛋白质2个, 上调表达蛋白质5个, 下调表达蛋白质1个。在9个蛋白质中, 有5个蛋白点得到鉴定。包括3个globulin-1 S allele precursor、1个2-isopropylmalate synthase B和1个translationally-controlled tumor protein。种子休眠破除过程中的蛋白质的变化说明种子经历了一系列生理生化活动, 深入研究这些蛋白质的生物学功能将有助于更清楚地认识玉米种子休眠问题。     

小麦蛋白质含量和优质亚基遗传

研究了小麦子粒蛋白质含量和优质高分子量麦谷蛋白亚基对小麦品质的影响及其互作关系。结果表明，控制蛋白质含量的基因作用主要是以加性效应为主，也存在非加性效应，F1的蛋白质含量与双亲的蛋白质含量的平均值高度相关，蛋白质含量的一般配合力方差大于特殊配合力方差，F2子粒蛋白质含量的分离呈正态分布。在优质育种亲本选配上，一般配合力比特殊配合力更重要一些。普通小麦品种高分子量麦谷蛋白亚基受遗传控制，不受环境影响，具有品种特性；其在F1中呈共显性和倾母遗传现象，在F2中遗传行为遵从孟德尔的基因独立分配和自由组合规律。在小麦品质改良中既要重视蛋白质的含量，更应重视蛋白质的质量，即数量和质量改良途径并重。     

玉米杂交种与亲本雌穗花器官形成期蛋白差异表达谱分析

为探讨玉米穗粒数杂种优势形成的分子机制,以玉米强优势杂交种豫玉22及其亲本综3和87-1的花器官形成期雌穗为材料,采用高通量的2-DE和MALDI TOF MS技术,建立了它们的蛋白差异表达谱,并对差异蛋白进行了质谱鉴定。结果显示,在检测到的1 290个蛋白点中,有114个(8.84%)在杂交种与亲本之间的表达差异达到显著水平,其中表现单亲沉默、偏高亲、中亲表达、偏低亲、杂种上调、杂种下调、亲本特异和杂种特异表达模式的蛋白点分别为27、25、15、13、11、11、10和2个。另外,成功鉴定出其中的104个差异表达蛋白点,涉及代谢、信号转导、能量、转录、蛋白质合成、蛋白运输与储藏、细胞生长与分裂、细胞结构、抗病防御、次生代谢、转座子及功能未知和假定蛋白等12个功能类别。玉米杂交种与亲本蛋白在丰度上的明显差异及涉及多个功能类别,可能与玉米穗粒数杂种优势的形成有关。     

Differential gene expression between wheat hybrids and their parental inbreds in seedling leaves

Differential display of mRNA was used to analyze the differences of gene expression in seedling leaves between heterotic hybrid/nonheterotic hybrid and their parental inbreds in order to study the molecular basis of heterosis in wheat. The results indicated that patterns of gene expression in hybrids differ significantly from their parents. Both quantitative and qualitative differences were observed. The quantitative differences include gene over-expression, gene under-expression in hybrid and dominant expression of highly-expressed parental genes in hybrids. The qualitative differences include silencing in hybrids of genes expressed either in male or female parent, and silencing in hybrids of genes expressed in both parents. Expression in hybrid of genes only expressed either in male or female parent was also observed. It was also found that some genes expressed at high level in heterotic hybrid were underexpressed or expressed at low level in nonheterotic hybrid. One differentially expressed cDNA fragment 4B was cloned and sequenced after being confirmed through Northern blot analysis. Homology search in GenBank proved that the cDNA fragment is a new sequence. The selection of primers for differential RNA display in wheat and the relationship between wheat heterosis and alteration of gene expression in hybrids as compared to their parental inbreds were also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

抗虫杂交棉F1代与亲本Bt蛋白表达量及抗虫差异性研究

对转 Bt基因抗虫杂交棉正、反交 F1代与抗虫亲本的 Bt蛋白表达量及抗棉铃虫差异性研究表明 :抗虫亲本与其杂种 F1代均高抗棉铃虫 ,但抗虫亲本的抗虫性略好于其杂种 F1代 ,并且明显地高于非抗虫亲本 ;正、反交杂种 F1代间的抗虫性几乎没有差异。生长前期的抗虫性好于后期 ,同一时期嫩叶或侧枝生长点的抗虫性好于幼蕾。抗虫亲本叶片和花瓣的 Bt蛋白含量明显地高于其杂种 F1代 ,抗虫亲本功能叶的 Bt蛋白含量明显地高于其上部非功能叶 ,而杂种 F1代功能叶的 Bt蛋白含量则明显地低于其上部非功能叶。盛花期后至吐絮期前 ,叶片和花瓣的 Bt蛋白表达量明显增加 ,在抗虫亲本中表现最为明显。与叶片相比 ,在花瓣中检测到的 Bt蛋白含量极低。正、反交 F1代间的 Bt蛋白表达量差异较小或无规律可循     

Relationship of genetic distance and hybrid performance in hybrids derived from a new photoperiod-thermo sensitive male sterile wheat line 337S

Heterosis has been exploited in many crops and made a significant contribution to the world food supply. Genetic distance (GD) is one of valuable criteria for selecting parents in hybrid breeding. The objectives of this study were to estimate GD between a novel photoperiod-thermo sensitive male sterile wheat line 337S and several common cultivars using simple sequence repeat (SSR) markers, and to evaluate the relationship of GD and heterosis. The line 337S, as a maternal parent, was crossed with 16 common wheat varieties to produce 16 hybrids. Eight agronomic traits were investigated for the parents and F1 hybrids. Mid-parent heterosis (MPH) and specific combining ability (SCA) were calculated. GD and cluster analysis were performed among the 17 parents. The correlations of GD with F1 performance, MPH and SCA were analyzed. The results showed that GD estimates among the 17 parents averaged 0.42 with a range from 0.26 to 0.57, and the averaged GD between 337S and the 16 parental cultivars was 0.44. Cluster analysis separated the 17 parents into three main groups. SSR markers were useful in analyzing genetic divergences among the 17 parental lines. However, GD based on SSR markers poorly correlated with F1 performance, MPH and SCA. Thus, GD revealed by SSR markers can not provide a reliable prediction in practical wheat hybrid breeding.     

玉米杂交种先玉335及其亲本种子萌发过程中胚芽蛋白质组学分析

以培养84 h种子的胚芽(包括胚芽鞘)为试验材料, 对优良玉米杂交种先玉335与其亲本自交系的蛋白质组学差异进行了分析。结果表明, 在杂交种先玉335中共检测到560个蛋白点, 在亲本PH6WC和PH4CV分别检测到507个和508个蛋白点, 而且先玉335胚芽及胚芽鞘中81%的蛋白点表现非加性累积模式, 其中 288个蛋白点表现为超高亲的上调表达, 仅15个蛋白点表现超低亲的下调表达, 因此推测非加性蛋白的累积可能是杂交种先玉335胚芽萌发过程中胚芽及胚芽鞘杂种优势产生的主要原因。用于质谱分析的13个差异极显著蛋白主要涉及代谢途径、蛋白折叠、胁迫响应、细胞骨架和细胞解毒5种类型。     

The genotypic and phenotypic interaction of wheat and rye storage proteins in primary triticale

The creation of new triticale germplasm from its wheat and rye progenitors is an important source of potentially beneficial alleles. This research focused on the inheritance of glutenin and secalin alleles in triticale and their interaction in the hybrid offspring in terms of both subunit expression and gluten strength. Multiple crosses between five durum lines and two inbred rye lines were performed to create a set of 23 fertile amphidiploid lines. The SDS-PAGE banding patterns of the denatured HMW glutenins, LMW glutenins, HMW secalins and 75k γ-secalins (termed secaloglutenin) were compared to their exact parental plants, and their SDS-sedimentation was evaluated. Secaloglutenin subunits were simply inherited and expressed in all primary triticales. The few observed variations in banding patterns were accounted for within measured heterogeneity of the exact rye parental plants, for which the secalin alleles were thoroughly described. However, the possibility remains that mixed protein oligomers were formed between polymeric and monomeric storage proteins at the ultrastructure level. Furthermore, significant differences in the gluten strength of primaries derived from different durum parents were observed. This research suggests triticale of high gluten strength can be obtained by selection of parents with favourable glutenin alleles.     

普通小麦品种间以及与六倍体小黑麦种间杂种F1农艺性状的遗传分析

为给杂交选育优良小麦品种提供基础,本研究以普通小麦品种晋农190为父本,分别与9个普通小麦品种、7个六倍体小黑麦品种为母本配置杂交组合,对杂种F1代的农艺性状进行分析,发现除了无芒对有芒为显性之外,其余农艺性状均表现为数量遗传,F1代表现为超亲遗传或介于双亲之间。六倍体小黑麦的灰绿色叶片或茎秆、穗长长和红色不饱满的籽粒在杂种F1代显现,可作为鉴别真假杂种的典型性状。     

Searching for DNA introgressed from wheat and for wheat-like grain proteins in a rice x wheat hybridization derivative

The DNA of a putative rice x wheat hybridization derivative (X Oryticum oryzoides) from China, the DNA of its parental rice cultivar and the DNA of a wheat line were digested with ten different restriction endonucleases, resolved by agarose electrophoresis, Southern blotted and hybridized using genomic wheat DNA as a probe. Phenol extracted, ethanol and cetyltrimethylammonium bromide precipitated DNA of the putative hybrid showed a restriction fragment length polymorphism (RFLP) different from that of the parental rice. When the DNA was further purified by Qiagen chromatography, the RFLP differences were not detected. Hence the apparent RFLP differences were probably due partial digestion of the less pure DNA preparations by the restriction endonucleases. No real introgressed fragments from wheat genome could be shown. The HpaII/MspI sites were more frequently digested with MspI than with HpaII in rice and hybridization derivative DNA, but the sites were evidently more frequently methylated in wheat DNA. Thus, in terms of methylation of the DNA, the hybridization derivative was much more like the rice parent than the wheat parent. The hybridization derivative showed a single endospermal protein (mass 19 kg mol^-1) not detected in the parental rice cultivar. This minor protein was soluble in buffered 50% isopropanol and precipitable with methanol. The results indicate that there are no or only short introgressed sequences from wheat in the rice/wheat derivative, a result which might be considered in breeding efforts with the hybrid derivative.     

过氧化物酶及种子蛋白分析鉴定甜椒杂交种纯度

用聚丙烯酰胺均匀胶和等电聚焦电泳技术，分析了甜椒种子及种苗的过氧化物同功酶谱和种子水溶蛋白图谱。在过氧化物酶谱带的两个位点ＰＲＸ－１和ＰＲＸ－２上，自交系亲本与Ｆ１杂交种有明显的差异。水溶蛋白的一个位点ＰＲＯＴ亲本与Ｆ１有明显差异。供试甜椒组合的过氧化物酶和蛋白的图谱，分别具有这些位点。     

利用RAPD标记鉴定大白菜杂交种纯度的研究

应用随机扩增多态性ＤＮＡ（ＲＡＰＤ）标记鉴定大白菜杂种商品种子的纯度。用５０个随机引物检测了２个杂交种北京５７号和北京１０６号及其亲本,引物ＯＰＥ－２０在北京５７号杂交种中产生了有别于双亲的特殊标记,引物ＯＰＨ－０６及ＯＰＨ－０７在北京１０６杂交种中产生了有别于双亲的特殊标记,能清楚地区分杂交种及其双亲,并将这些引物应用在北京５７号和北京１０６号杂交种样品的纯度检测中。这个结果显示了ＲＡＰＤ标记在大白菜杂交种商品种子纯度检测上的实际用途。