枇杷试管苗2个CAT基因成员的克隆及表达分析

以早钟6号枇杷试管苗叶片为试验材料,采用同源克隆方法从叶片mRNA中分离得到CAT基因的2个成员Ej-CAT1和Ej-CAT2,Ej-CAT1 cDNA序列全长1 738 bp,包括1 479 bp ORF和257 bp 3′-UTR;Ej-CAT2序列全长1 742 bp,包含1 479 bp ORF及261 bp 3′-UTR。2个CAT基因成员的核苷酸序列相似性为98.04%,其开放阅读框推导的氨基酸序列具有97.32%的相似性。生物信息学分析结果表明:2个蛋白均编码492个氨基酸,是亲水蛋白,不含信号肽,不存在螺旋卷曲结构,是非跨膜蛋白,亚细胞定位于过氧化物酶体;Ej-CAT1及Ej-CAT2蛋白分别有18、17个功能位点,22、19个磷酸化位点。CAT基因在枇杷试管苗离体保存过程中不同时期具有不同的表达量,整体趋势呈"W"形状,在离体保存1个月时表达量最高,保存7个月时表达量最低。CAT表达量变化可能跟枇杷试管苗的生长发育模式有关。     

孔雀草试管苗叶片AP1基因的克隆及其生物信息学分析

以孔雀草试管苗叶片为材料,成功克隆了孔雀草AP1基因的cDNA全长,并对其进行了生物信息学分析。AP1基因全长919 bp(GenBank登录号为JX310277),其中5′-UTR 92bp、3′-UTR 149bp、3′端poly(A)尾巴25 bp,开放阅读框为678 bp,编码225个氨基酸。AP1-1可能存在于细胞核中,为亲水蛋白,不含信号肽,共形成4个卷曲螺旋结构;二级结构主要有α螺旋和无规则卷曲构成,存在亮氨酸拉链结构和1个MADS-box区。此蛋白可能发生磷酸化位点的位置有7个。从系统进化树分析表明,该蛋白与甘菊具有较高的亲缘关系。     

金弹试管苗叶片MLP2基因的克隆及其生物信息学分析

采用RT-PCR结合RACE技术,以金弹(尤溪金柑Fortunella crassifolia cv.Youxijingan)试管苗的叶片为材料,克隆得到金弹Fc-MLP2基因的全长cDNA序列,其在GenBank中的登录号为JX310276.Fc-MLP2基因全长908 bp,含有一个669 bp的开放阅读框,编码223个氨基酸.生物信息学分析表明:该蛋白是定位于细胞外的一种疏水性分泌蛋白,具有2个功能位点、13个磷酸化位点和信号肽,其二级结构由α螺旋、β折叠、β转角、无规则卷曲组成.系统进化树分析表明,金弹Fc-MLP2与粗皮柠檬的RlemMLP2蛋白亲缘关系较近,聚为同一类.该蛋白可能在植物抗虫、抗病原菌、抗饥饿和延缓植物衰老方面起到一定作用.     

文心兰OnCOBRA基因克隆及表达模式分析

以文心兰试管苗为材料,采用RT-PCR结合RACE法,克隆文心兰OnCOBRA基因的cDNA全长和DNA序列。结果表明:COBRA全长为1 601 bp,开放阅读框(ORF)为1 386 bp,共编码461个氨基酸;OnCOBRA的DNA序列共2 949 bp,且含有6个外显子和5个内含子。生物信息学结果表明,OnCOBRA属于不稳定的疏水蛋白,具有信号肽、跨膜结构和CCVS保守区域,亚细胞定位于细胞膜中;与无油樟、玉米、籼稻、拟南芥等具有较高的同源性。系统进化树分析结果表明,文心兰OnCOBRA蛋白与玉米(ZmCOBRA)、无油樟(AtCOBRA)、籼稻(OsCOBRA)处于统一分枝,推测OnCOBRA基因是COBRA基因家族的成员。qPCR结果表明,OnCORBA为组成型表达,在成苗期表达量最高,在文心兰类原球茎时期表达量最低。     

玉米转录因子Zmhdz6的表达模式分析及互作功能蛋白预测

利用RT-PCR方法从玉米基因组中克隆到GRMZM2G056600(Zmhdz6)转录因子的cDNA序列长786 bp,编码261个氨基酸,分子质量为28.46 kD。二级结构显示,编码的蛋白主要以α-螺旋和不规则卷曲为主。亚细胞定位预测显示,Zmhdz6蛋白定位于细胞核。进化树和motifs分析发现,Zmhdz6基因属于HD-Zip-I家族成员且与高粱同源达到99%,单子叶植物亚群含有7个重要的motifs。Zmhdz6蛋白互作预测,发现互作的基因主要参与逆境胁迫、信号传导和植物生长发育过程。荧光定量结果显示,Zmhdz6基因在雌穗和雄穗中高度表达,NaCl、PEG胁迫和外源ABA诱导时均上调表达。结果说明Zmhdz6基因可能参与玉米逆境胁迫的应答和信号传导路径。     

甘蔗可溶性酸性转化酶基因的克隆及序列分析

可溶性酸性转化酶在植物糖代谢中起着关键作用,为了研究甘蔗中可溶性酸性转化酶,利用电子克隆与RT-PCR方法从甘蔗中分离出1个甘蔗可溶性酸性转化酶基因DNA及cDNA序列,DNA序列长度为3 266 bp,含有3个外显子和2个内含子,cDNA序列长度为1 890 bp,包含1 656 bp的完整开放阅读框,编码551个氨基酸。BLASTX分析显示,该蛋白与其它植物已知的酸性可溶性蔗糖转化酶具有很高的同源性,推测该蛋白为可溶性酸性转化酶。预测甘蔗SAI家族可能只有1个基因,这可能与其大量积累蔗糖的能力有重要关系。     

巴西橡胶树中碱性转化酶HbNINa基因的克隆和表达模式分析

检索橡胶树EST和基因组数据库,发现了1个在叶片中高丰度表达的中碱性转化酶基因.采用RT-PCR和RACE技术获得了该基因的cDNA和基因组序列,命名为Hb NINa.序列分析结果显示,HbNINa基因cDNA序列的编码区(CDS)序列长度为1 719 bp,对应的基因组序列为3 696 bp.序列分析表明,该基因编码571个氨基酸多肽,推测其分子量大小为65.7 ku,等电点为6.36.HbNINa蛋白包含植物中碱性转化酶的全部保守结构域.基因组结构分析显示Hb NINa基因包含4个外显子和3个内含子.QPCR分析结果表明,HbNINa基因在在胶乳中表达水平极低,而在其他组织如树皮、树叶和雌花中的表达丰度相对较高.在叶片发育过程中,Hb NINa在叶片发育的初期高丰度表达,推测HbNINa可能参与叶片蔗糖利用,进而在橡胶树叶片发育过程中起重要作用.     

TaMBD2基因cDNA全长克隆及其在小麦叶片和种子中的表达

为探讨甲基结合蛋白(MBD)在小麦生长发育过程中的生物学功能,通过RACE技术克隆了小麦TaMBD2基因的cDNA全长,并分析了该基因在小麦叶片、种子发育及萌发过程中的表达特性。RACE克隆及序列分析表明,TaMBD2基因cDNA全长为1 511 bp,其中5 UTR 164 bp,3 UTR 405 bp,ORF 942 bp。对该基因编码氨基酸序列的分析发现,TaMBD2编码蛋白中包含有1个典型的甲基结合域和1个CW型锌指结构域;通过RT-PCR分析发现,TaMBD2基因在叶片中的表达随着小麦的生长发育而逐渐增强;在种子发育过程中,该基因在花后20和30 d时的表达量最高;在种子萌发过程的胚和胚乳中,该基因的表达水平变化不大。     

早钟6号枇杷两个ETR基因成员的克隆及序列分析

以早钟6号枇杷的试管苗叶片为材料,采用同源克隆方法得到乙烯受体基因ETR的两个成员Ej-ETR1a和Ej-ETR2a的cDNA全长序列,在GenBank上的登录号为JX307084和JX307089.Ej-ETR1a序列全长2771 bp,包括227bp 5’-UTR、2226 bp ORF及318 bp 3’-UTR; Ej-ETR2a序列全长2594 bp,包含2226 bp ORF及366 bp 3’-UTR.Ej-ETR1a、Ej-ETR2a均编码741个氨基酸,且氨基酸序列具有95.28％的相似性.生物学信息分析表明:枇杷Ej-ETR1a和Ej-ETR2a均属于乙烯受体ETR1亚家族,为一个具有跨膜结构的非分泌蛋白,具有疏水性.2个ETR成员的克隆为进一步研究枇杷乙烯受体作用机制奠定了基础.     

茶树CsASR基因的克隆及其表达分析

逆境胁迫影响茶树生长发育及茶叶品质,ASR(abscisic acid,stress,ripening-induced)基因在植物抗逆响应中具有重要功能。本研究以龙井43品种茶树为材料,从中克隆了CsASR基因的全长cDNA序列、基因组序列及其启动子序列,分析了该基因的生物信息学特征及在组织间和不同胁迫处理下的表达模式。结果显示,CsASR的cDNA序列全长875 bp,含有546 bp的ORF序列,编码181个氨基酸,蛋白质分子量19.89 kD,理论等电点5.69;CsASR蛋白结构序列中74.5%的序列为无序结构,是一种无序蛋白;CsASR的C-端含有ABA/WDS功能结构域,主要定位于细胞质和细胞核中;茶树CsASR与海枣的ASR相似性最高,为87%,而在进化树中与枣的关系最近。CsASR基因含2个外显子,第1个外显子长363 bp,第2个外显子长183 bp,内含子较大为2 750 bp,内含子中含7种简单重复序列和2种DNA转座子序列。克隆获得起始密码子ATG上游2 554 bp的启动子区序列,该启动子上含有干旱、低温、高温以及ABA等相关的顺式作用元件。CsASR在根中的表达量最低;ABA抑制CsASR的表达,而干旱、NaCl和低温胁迫能够显著上调CsASR的表达。表明CsASR基因可能与茶树抗逆密切相关。     

Synthesis and thermo-responsive properties of chitosan-g-poly (N-isopropylacrylamide) and HTCC-g-poly(N-isopropylacrylamide) copolymers

Carboxyl group-terminated poly(N-isopropylacrylamide) (PNIA-COOH) was synthesized via radical polymerization of N-isopropylacrylamide (NIA) using mercaptoacetic acid (MAA) as a chain transfer agent. The molecular weight of the PNIA-COOH was controlled by changing the molar ratio of MAA to NIA. A water-soluble chitosan derivative, N-(2-hydroxy)propyl-3-trimethylammonium chitosan chloride (HTCC), was also synthesized by reacting chitosan with glycidyltrimethylammonium chloride. Then, chitosan-g-PNIA and HTCC-g-PNIA copolymers were synthesized using the “graft-onto” method by reacting PNIA-COOH with chitosan and HTCC, respectively. The formation of the grafted copolymers was confirmed by Fourier transform infrared spectroscopy, solubility test in water, and scanning electron microscopy — energy dispersive spectroscopy. The thermo-responsive behaviors of the grafted copolymers and the change in lower critical solution temperature (LCST) were also studied. Chitosan-g-PNIA was insoluble in water and behaved like a thermo-responsive hydrogel due to the crosslinking-point action of the chitosan backbone. The swelling ratio of chitosan-g-PNIA increased with increasing PNIA content. HTCC-g-PNIA behaved as a water-soluble thermo-responsive polymer. Compared to the homo PNIA, the LCST of HTCC-g-PNIA was slightly increased.     

Spinodal phase separation and isothermal crystallization behavior in blends of VDF/TrFE(75/25) copolymer and poly(1,4-butylene adipate) (I)

Phase behavior and spinodal phase separation kinetics in binary blends of a random copolymer of vinylidene fluoride and trifluoroethylene (75/25) [P(VDF/TrFE)] and poly(1,4-butylene adipate) (PBA) have been investigated by means of optical microscopic observation and time-resolved light scattering. The blends exhibited a typical lower critical solution temperature (LCST)∼34 °C above the melting temperature of the P(VDF/TrFE) crystals over the entire blend composition range. P(VDF/TrFE) and PBA were totally miscible in the temperature gap between the melting point of P(VDF/TrFE) and the LCST. Temperature jump experiments of the 3/7 P(VDF/TrFE)/PBA blend were carried out on a light-scattering apparatus from a single-phase melt state (180 °C) to a two-phase region (205∼215 °C). Since the late stage of spinodal decomposition (SD) is prevalent in the 3/7 blend, SD was analyzed using a power law scheme. Self-similarity was preserved well in the late stage of SD in the 3/7 blend.     

花生油酸亚油酸比值（O/L）的遗传分析

应用植物数量性状主基因+多基因混合遗传模型，对花生品种“青苗豆×全州麻壳”、 “富川大花生×隆安宝湾花生”和“汕油162×Sunoleic95R”三个不同杂交组合的F2分离群体的油酸/亚油酸比值进行了单个分离世代的数量性状分离分析，结果表明：“青苗豆×全州麻壳”、“富川大花生×隆安宝湾花生”两个杂交组合结果一致，O/L受一对负向完全显性的主基因控制，F2群体表现为由主基因型AA和Aa+aa按1：3比例的混合；另一个组合“汕油162×Sunoleic95R”的油酸/亚油酸比值受两对加性-显性-上位性主基因控制遗传，两对主基因间的基因效应差异不大。第一对主基因加性效应（da）1.3586和第二对主基因加性效应（db）1.3580几乎相同。三个组合的主基因遗传力分别为61.27﹪、64.09﹪和64.66﹪，多基因的遗传力分别为33.85％、34.52％和33.45％。     

花生油酸/亚油酸比值(O/L)遗传研究

研究采用三个不同的杂交组合,"青苗豆×泉州麻壳"、"富川大花生×隆安宝湾花生"和"汕油162×Sunoleic95R",分别对其F2代油酸/亚油酸比值(O/L)进行柱形图分析.结果表明,不同的亲本其后代呈现出不同的图形分布,一个杂交组合后代呈现多峰,另外两个是偏离正态分布的单峰,根据图形分布可以判断出O/L是由主基因控...     

Discrimination of sterile oat (Avena sterilis) in winter barley (Hordeum vulgare) using QuickBird satellite images

Site-specific weed management implies detecting the location of weeds in order to generate maps of their spatial distribution. This information facilitates a more accurate application of herbicides, spraying them in the exact areas of weed growth and in the required doses. In order to explore the potential of commercial satellites to discriminate and map weeds, we used the information contained in high spatial resolution images acquired by the QuickBird satellite to assess the density of sterile oat (Avena sterilis) present in a winter barley field at two different dates (March and June). Our results confirmed the potential of using satellite images in the spectral discrimination of weed patches in infested fields. The results of binary logistic regressions showed that the best matches in the classification of three categories (low, medium, or high sterile oat densities) corresponded to the March image. QuickBird’s March image provided reliable estimates of sterile oat patches in barley crops when weed density was relatively high (between 86% and 94% of agreement between predicted and observed densities). However, when weed densities were lower than 10 plants/m² there were serious difficulties to distinguish them from weed-free zones (between 72 and 75% of global agreement in the classification) with large underestimation of medium density weed patches (10 plants/m²). This is a potential limitation considering than the thresholds used for herbicide application decisions are generally close to this density. However, the information obtained may still be useful for producing field maps to describe the spatial distribution of this weed. Moreover, these studies have provided valuable information on the best spectral regions and/or vegetation indices for approaching discrimination between sterile oat and cereal crops and the most suitable period for it.     

Pest-damage relationships for Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) on soybean (Glycine max) and dry bean (Phaseolus vulgaris) during pod-fill

The response of soybean (Glycine max) and dry bean (Phaseolus vulgaris) to feeding by Helicoverpa armigera during the pod-fill stage was studied in irrigated field cages over three seasons to determine the relationship between larval density and yield loss, and to develop economic injury levels. H. armigera intensity was calculated in Helicoverpa injury equivalent (HIE) units, where 1 HIE was the consumption of one larva from the start of the infestation period to pupation. In the dry bean experiment, yield loss occurred at a rate 6.00 ± 1.29 g/HIE while the rates of loss in the three soybean experiments were 4.39 ± 0.96 g/HIE, 3.70 ± 1.21 g/HIE and 2.12 ± 0.71 g/HIE. These three slopes were not statistically different (P > 0.05) and the pooled estimate of the rate of yield loss was 3.21 ± 0.55 g/HIE. The first soybean experiment also showed a split-line form of damage curve with a rate of yield loss of 26.27 ± 2.92 g/HIE beyond 8.0 HIE and a rapid decline to zero yield. In dry bean, H. armigera feeding reduced total and undamaged pod numbers by 4.10 ± 1.18 pods/HIE and 12.88 ± 1.57 pods/HIE respectively, while undamaged seed numbers were reduced by 35.64 ± 7.25 seeds/HIE. In soybean, total pod numbers were not affected by H. armigera infestation (out to 8.23 HIE in Experiment 1) but seed numbers (in Experiments 1 and 2) and the number of seeds/pod (in all experiments) were adversely affected. Seed size increased with increases in H. armigera density in two of the three soybean experiments, indicating plant compensatory responses to H. armigera feeding. Analysis of canopy pod profiles indicated that loss of pods occurred from the top of the plant downwards, but with an increase in pod numbers close to the ground at higher pest densities as the plant attempted to compensate for damage. Based on these results, the economic injury levels for H. armigera on dry bean and soybean are approximately 0.74 HIE and 2.31 HIE/m², respectively (0.67 and 2.1 HIE/row-m for 91 cm rows).     

Discrimination of weed seedlings, wheat (Triticum aestivum) stubble and sunflower (Helianthus annuus) by near-infrared reflectance spectroscopy (NIRS)

The potential of using near-infrared reflectance spectroscopy (NIRS) to discriminate between seven broadleaf weed species, sunflower and wheat stubble was evaluated. Our results showed that the spectral window between 750 and 950 nm, which corresponds to near-infrared wavelength, was able to discriminate among wheat stubble, sunflower and the most problematic “hard-to-control” weeds in no-till sunflower, i.e. Little mallow (Malva parviflora L.) and Ecballium elaterium L. These results are promising for further work in real-time remote sensing identification of weed patches in sunflower fields.     

Apigenin di-C-glycosides (ACG) content and composition in grains of bread wheat (Triticum aestivum) and related species

Apigenin di-C-glycosides (ACGs) are present in the grain of bread wheat and other related cereals primarily as one or two sets of Wesseley-Moser isomers containing either arabinose and glucose (ACG1) or arabinose and galactose (ACG2) on the A ring of apigenin. These compounds may contribute to the yellow colour of wheat-based products made under alkaline conditions and in addition, have possible roles in a number of plant physiology processes and human health. The aims of this investigation were to survey genetic variation for ACG content and composition in hexaploid bread wheat (Triticum aestivum L.) and to examine ACGs in the putative progenitors of hexaploid wheat and available genetic stocks as a first step towards understanding the mechanisms involved in their biosynthesis and genetic control. Substantial variation in both grain ACG content and the ratio, ACG1/ACG2, were identified within bread wheat cultivars and related species. Genotype controlled the major portion of the variation. ACG content appeared to be a multigenic trait whereas variation in ACG1/ACG2 was associated with a limited number of chromosomes, in particular chromosomes 1B, 7B and 7D. The results suggest that it should be possible to manipulate both ACG content and composition traits through breeding.     

Effects of sulfosulfuron soil residues on barley (Hordeum vulgare), sunflower (Helianthus annuus) and common vetch (vicia sativa)

The phytotoxicity and sensitivity of succeeding crops to the new sulfonylurea, sulfosulfuron, have been reported although there is insufficient data on the phytotoxic effect of field soil residues. Growth chamber bioassays were conducted to detect the presence of residues in soil samples previously treated with sulfosulfuron at the recommended rate and double rate (20 and 40 g a.i./ha) that could affect the succeeding crop. Soil samples were collected between 7 and 9 months after sulfosulfuron application in field selectivity assays at nine different locations in Northern and Central Spain. The bioassay test species were barley (Hordeum vulgare L.) ‘Graphic’, sunflower (Helianthus annuus L.) ‘Albasol’ and common vetch (Vicia sativa L.) ‘Neska’, typical crops grown in rotation with wheat (Triticum aestivum L.) in this area. Sulfosulfuron residues did not affect barley and common vetch, but inhibited shoot length, root length and root dry weight of sunflower seeded into some soils treated with the 2× rate (40 g a.i/ha) 9 months earlier.     

An effect of rust (Uromyces viciae-fabae) on the yield of spring-sown field beans (Vicia faba) in the UK

Rust (Uromyces viciae-fabae) has recently been noted with increased frequency and earliness on spring-sown beans (Vicia faba) at Rothamsted. In 1982 an experiment was made to assess the damage and yield loss, if any, being caused by the disease. Plots received benomyl sprays to control chocolate spot (Botrytis fabae) or not, maneb plus mancozeb (protective) or propiconazole (systemic) fungicide applied twice or three times to control rust, or no fungicide at all. Rust was first found on 7 July and sprays were applied from 9 July but by 19 August plants of all treatments were dead. Fungicides controlled disease development during July and at harvest overall yields were 4·51 and 5·43 t/ha in unsprayed and sprayed treatments respectively. The difference was accounted for by the weight of individual grains and not by the number of pods per plant or grains per pod. Previously rust has been considered unimportant in the UK but its potential for damage is considerable.