花生中UDP-glucosyltransferase基因的克隆及在非生物胁迫下的表达研究

通过RACE技术从花生叶片cDNA中克隆到了UDP-glucosyltransferase基因,命名为AhUGT83A1-like （Genbank注册号为KF411463）。该基因全长为1 530bp,ORF为1 380bp,编码460个氨基酸。序列比对与进化分析结果表明,该序列有保守的UDPGT结构域,并且与其它植物的UGT蛋白同源性较高。荧光定量PCR结果表明,AhUGT83A1-like基因在高盐和低温处理的花生根和叶片中均上调表达;在干旱处理时,根中表达也受到显著上调,但叶片中表达量则有明显下降。以上结果表明AhUGT83A1-like可能参与了花生对干旱、低温和高盐的抗性调控。ABA处理结果表明,AhUGT83A1-like在花生根中对ABA响应明显,但在叶片中对ABA没有明显响应,表明该基因在花生根中对非生物胁迫的调控可能是以依赖ABA的方式发挥作用的。?     

花生CONSTANS-Like（COL）家族基因的克隆与表达分析

开花是植物从营养生长到生殖生长的重要过程,CONSTANS-Like（COL）基因是植物光周期诱导开花途径中的关键转录因子,但是在花生中的具体功能尚不清楚。本研究通过生物信息学的方法对花生COL 基因家族成员序列特征、系统进化关系、基因结构、保守结构域及表达模式进行分析,最终克隆了17个AhCOL 基因（AhCOL1-Ah⁃COL17）。花生17个AhCOL 基因氨基酸长度范围为327aa~617aa,等电点（pI）均小于7,在蛋白质序列的氮端与碳端 均含有保守的BBX和CCT结构域。此外,不同组织中的基因表达模式分析,表明多数AhCOL 基因在叶片中的表达量显著高于其他组织。不同时期的表达量分析表明多数AhCOL 基因的表达量从苗期到开花期呈现递增。本研究为研究花生AhCOL 基因的潜在功能奠定了重要基础。     

花生蛋白质含量的主基因加多基因遗传分析

以郑8903×豫花4号构建的包含215个家系的重组自交系群体为材料,在海南三亚和河南原阳两个环境下种植,采用凯氏定氮仪测定蛋白质含量,运用数量性状主基因加多基因混合遗传模型分析方法,开展了花生蛋白质含量的遗传模型分析。结果表明,两个环境条件下家系间蛋白质含量均存在广泛变异,表现超亲遗传现象,其频数分布图呈正态分布特征。在两个环境中蛋白质含量的遗传均符合多基因遗传模型（C模型）,即受多基因效应和环境作用,其多基因遗传率分别为29.63%和18.77%;环境引起的变异分别为46.05%和54.08%。     

盐旱交叉胁迫对花生生长发育和生理特性的影响

为了探究盐旱交叉胁迫对花生生长发育的影响,以抗旱不耐盐花生品种花育22和抗旱耐盐花生品种花育25为试验材料,通过防雨棚盆栽试验研究了干旱、盐、盐+干旱、干旱后复水+盐等4种胁迫对花生产量、农艺性状、生物量、叶绿素SPAD值、丙二醛含量、活性氧清除能力及渗透调节物质含量的影响。结果显示：各胁迫处理均显著抑制了两个花生品种植株的生长和荚果产量,其中,盐胁迫对花育22生长的影响大于干旱胁迫;盐+旱胁迫下,两个花生品种受伤害程度最大,产量最低。与单一盐胁迫相比,干旱预处理提高了盐胁迫后期花生超氧化物歧化酶和抗坏血酸过氧化物酶活性,增强了植株活性氧清除能力,降低了叶片丙二醛含量,从而缓解盐胁迫对膜系统的过氧化伤害,提高了叶绿素含量,促进了植株生长,增加了干物质积累,最终提高盐胁迫下花生产量。另外,与单一盐或干旱胁迫相比,盐+旱胁迫对花育22和花育25的伤害均加重,而干旱预处理有利于2个品种在盐胁迫下活性氧代谢和光合色素的提高,促进植株的生长,提高植株对盐胁迫的交叉适应能力,从而缓解盐胁迫对花生植株的抑制作用。      

渗透胁迫对花生幼叶活性氧伤害和膜脂过氧化作用的影响

对３种不同抗旱性的花生栽培品种进行渗透胁迫处理。结果表明,渗透胁迫使花生幼叶水势（ΨＬ）下降,花生幼叶的Ｏ２、产生速率随渗透胁迫处理强度加大而增加,ＭＤＡ含量以及ＳＯＤ、ＰＯＤ和ＣＡＴ的活性水平变化与Ｏ２产生速率的变化趋势相似。ＭＤＡ含量明显增高,其过程与不势呈负相关与ＲＰＭＰ呈正相关,均达极显著水平,渗透胁迫处理过程中,ＧＳＨ和ＡＳＡ含量下降。与泉花１０号和汕油７１相比白皮１号ＲＰＭＰ增加幅度、     

花生硫氧还蛋白AhTRX h基因的克隆及表达分析

h型硫氧还蛋白(TRX h)是最多的一类硫氧还蛋白,在种子萌发、氧化还原、信号传递以及响应生物和非生物胁迫等多种途径中起重要作用。以花生铝敏感品种‘中花2号’和耐铝品种‘99-1507’为材料,对h型硫氧还蛋白亚族成员AhTRX h进行克隆和表达分析。结果表明:该基因包含长为429 bp的完整开放阅读框,编码142个氨基酸;进化树分析表明,花生TRX h与鹰嘴豆TRX h、拟南芥TH9亲缘关系较近,且含有保守活性位点WCGPC;亚细胞定位显示,AhTRX h定位于叶绿体;构建原核表达重组质粒pGEX-6p-1-AhTRX h,转化大肠杆菌Rosetta菌种,在上清和沉淀均有表达,纯化获得可溶的具活性的重组蛋白,体外检测重组蛋白没有明显的发生亚硝基化的趋势。酵母双杂显示,AhTRX h蛋白没有自激活性,筛选花生文库获得互作蛋白——钙网蛋白和金属硫蛋白。该结果表明AhTRX h可能通过与钙网蛋白和金属硫蛋白相互作用,参与花生对铝胁迫响应。     

花生AhARF 基因家族鉴定与表达分析

生长素响应因子（Auxin response factor,ARF）是生长素信号转导途径中的一类重要转录因子,在种子萌发、器官形成、果实成熟、胚胎发生等多个生长发育过程发挥作用。为了揭示花生基因组中AhARF 基因家族的特征,本研究利用生物信息学方法从花生基因组中鉴定了62个花生AhARF 家族基因,它们不均匀分布于除1号和11号之外的其他18条染色体上,在A和B亚基因组对应关系的染色体上数目大体相同。依据系统发生关系,62个花生AhARFs 与拟南芥AtARFs 家族基因共同聚在除ClassIII（仅包含拟南芥AtARFs）外的其余4个分支。共线性分析共检测到33对片段重复基因,其Ka/Ks比值均小于1,受到环境压力的纯化选择。本研究还分析了它们的理化性质、蛋白保守结构域等特征。此外,基于22个组织转录组数据,绘制了62个花生AhARF 家族基因的表达模式热图。进一步应用qRT-PCR方法,分析了可能与萌发相关的4个AhARF 基因（AhARF10、AhARF20、AhARF23 和AhARF46）的时空表达模式。本研究可为种子萌发相关AhARF 基因的挖掘与功能鉴定提供基础。     

花生白藜芦醇和查尔酮合成酶基因的鉴定与表达分析

花生含有丰富的白藜芦醇,具有很好的营养价值和保健功能。白藜芦醇合酶（STS）是白藜芦醇合成途径中的关键酶。本研究运用生物信息学方法,从花生基因组数据中鉴定出了50个STS基因,17个编码查尔酮合成酶CHS基因;两者同属聚酮化合物合成酶家族。对所鉴定基因组家族成员的染色体定位、系统发育、外显子-内含子结构、基因表达模式分析,结果表明大量STS基因在根、种皮、果皮、果针中高量表达,45个STS和8个CHS基因均应答紫外胁迫。本研究结果为花生STS/CHS基因家族成员的功能鉴定提供有价值的参考信息,为花生的品质改良提供理论依据与基因资源。     

花生紫色酸性磷酸酶基因家族的鉴定及表达分析

紫色酸性磷酸酶（purple acid phosphatase, PAP）是一类金属磷酸酯酶,参与植物的磷素利用、碳代谢、细胞壁合成等多种生理功能,尤其是对磷缺乏的适应。本研究利用生物信息学手段在花生（Arachis hypogaea L.）全基因组水平上对PAP基因家族进行了鉴定,并对其系统进化关系、保守结构域、基因结构以及它们在22个组织中的表达模式进行了分析。结果表明：花生基因组中有39个AhPAP基因,氨基酸序列长度为205~905,等电点大多数都小于7,蛋白质C端均含有金属磷酸酶结构域。以花生、拟南芥、水稻、苜蓿共74个PAP蛋白构建的系统发育树可分为四组,每组中都含有来自不同物种的PAP,并没有因为物种差异而各自聚为一类。AhPAP基因家族中部分成员的表达具有组织特异性,arahy.P03NME、arahy.DAPS6C在根瘤中表达量最高,而它们在其他组织中表达量较低或未检测到表达。本研究为揭示AhPAP基因家族在花生中的生物学功能奠定了基础。     

Cold tolerance during early reproductive growth of chickpea (Cicer arietinum L.): genetic variation in gamete development and function

Chilling temperatures at flowering cause floral abortion in most chickpea cultivars. Recent evaluations of germplasm showed distinct genotypic differences in pod and seed set at low temperature but the morpho-physiological basis for such variation is unclear. Observations in the field during cold spells of December and January, and at 15/5°C and 15/0°C (day/night) regimes in growth rooms showed distinct genetic variation in flower morphology, gamete development (viability and size of pollen and ovules) and function (pollen germination and tube growth, ovule viability and fertilization, etc.). The greater pod-setting ability of tolerant lines (ICCVs 88502 and 88503) than the sensitive cultivars (Annigeri, Pant G 114, etc.) was associated with a higher pollen vigour (germination and tube growth) and ovule viability at low temperature in the former. The number of ovules was not affected by cold stress in all cultivars/lines but pollen size and viability were reduced in Annigeri. The reduced ovule fertilization, associated with decline in pollen tube growth and ovule viability, was the major cause for poor seed set at low temperatures. The magnitude of effects on gamete function varied with cultivar/line and severity of stress. Function of both pollen and ovules was adversely affected in Annigeri in 15/5° and 15/0°C regimes. In contrast, the reduction in pollen vigour was more than in ovule viability in ICCV 88510 and Pant G 114 at 15/5°C. At 15/0°C, however, both pollen vigour and ovule viability were reduced. A small increase in pod set of sensitive genotypes from manual pollinations with pollen from plants in the warm regime further suggested that pollen function was more adversely affected than pistil function at moderately low temperatures.     

Root development and the expression of aquaporin genes in rice seedlings under osmotic stress

We conducted hydroponic culture experiments to characterize root traits in a rice cultivar ‘Puluik Arang’ that has been identified in a previous study as a cultivar that is adaptable to unflooded conditions. Root morphological traits and the expression of 11 aquaporin genes in rice seedlings (cv. Puluik Arang and cv. Akitakomachi) subjected to osmotic stress by polyethylene glycol (PEG) treatments (10 and 20%) were analysed. ‘Puluik Arang’ exhibited significantly greater water uptake under 10% PEG treatment than ‘Akitakomachi’. Lateral root development was maintained in ‘Puluik Arang’ under PEG treatments. The expression of some aquaporin genes, particularly OsTIP2;1, was higher in ‘Puluik Arang’ than in ‘Akitakomachi’. Immunocytochemical analysis showed that the OsTIP2;1 protein mainly accumulated in endodermal cells. The results suggest that better lateral root development and the function of aquaporins could contribute to water uptake in ‘Puluik Arang’ under osmotic stress.     

In vitro and in silico analysis of two genes (A2-1 and G1-1) differentially regulated during dormancy and sprouting in potato tubers

Summary Two cDNAs, corresponding to genes differentially regulated during dormancy and sprouting in potato tubers, cultivar Désirée, were isolated: i) G1-1 corresponded to a gene that was turned off during dormancy and turned on during early phases of sprouting; ii) A2-1 corresponded to a gene activated during dormancy and strongly repressed during the transition from dormancy to sprouting. When induced, both genes were expressed at low level. Full-length cDNAs and genomic clones were isolated and characterized. G1-1 was a short gene, 452 bp long, containing an intronless open reading frame, coding for a putative protein of 64 aminoacids. Sequence analysis showed that G1-1 was homologous to an expressed sequence tag (EST) ofArabidopsis thaliana. A2-1 full-length cDNA was 1577 bp long and contained an open reading frame coding for a putative protein of 383 aminoacids, which contained a Walker box binding domain, common to a multifunctional family of intracellular ATPases.     

土壤干旱胁迫对白芥光合器官发育及光合速率的影响

利用旱棚盆栽研究了在土壤干旱胁迫下白芥抗旱和不抗旱品系的单叶面积,单株叶面积、出叶速度、叶绿素含量和和净光合速率。结果表明,在土壤干旱胁迫下不同白品系在不同生育时期的单叶面积、单株叶面积、出叶速度及净光合速率都下降,在一定土壤水分范围内,叶绿素含量随土壤水分的下降而增加,抗旱性强的品系受土壤干旱胁迫的影响较小。     

Characterization and Developmental Expression Patterns of Four Hexamerin Genes in the Bumble Bee,Bombus terrestris (Hymenoptera: Apidae)

Hexamerins are members of the hemocyanin superfamily and play essential roles in providing amino acids and energy for the nonfeeding stages of insects. In this study, we cloned and analyzed the expression patterns of four hexamerin genes (hex 70a, hex 70b, hex 70c, and hex 110) at different worker development stages and queen diapause statuses in the bumble bee, Bombus terrestris. The results of this study showed that hex 110 has the longest open reading frame (ORF; 3,297 bp) compared to the ORFs of hex 70a (2,034 bp), hex 70b (2,067 bp), and hex 70c (2,055 bp). The putative translation product of Hex 70a, Hex 70b, Hex70c, and Hex 110 has 677, 688, 684, and 1,098aa with predicted molecular mass of 81.13, 79.69, 81.58, and 119 kDa. In the development stages of workers, the expression levels of hex 70a, hex 70b, and hex 70c increased gradually from the larval stage and exhibited high expression levels at the pink eyed and brown eyed pupae stage, whereas hex 110 exhibited the highest expression level at the larval period. Four hexamerin genes were highly expressed at the prediapause status of queen (P < 0.05), and compared to the eclosion queen, the lowest upregulation was 3.7-fold, and the highest upregulation was 1,742-fold. The expression levels of hex 70b, hex 70c, and hex 110 at diapause were significantly higher than those at postdiapause (P < 0.05). In conclusion, hexamerins may play important roles in queen diapause and metamorphosis of larval and pupal stages.     

油菜种子含油量相关的脂肪酶基因BnSDP1克隆与表达分析

通过电子延伸和RACE (cDNA末端快速扩增) 克隆得到一个与油菜种子含油量相关的脂肪酶基因,命名为BnSDP1,其开放阅读框为2 472bp,编码分子量为92kD的蛋白,预测等电点为6.75,具有保守的patatin结构域,跨膜域位于N端和中间,序列分析显示BnSDP1拥有GXSXG脂肪酶的基本特征序列GSSVG。利用RT-PCR技术分析发现,在种子发育的成熟期（即脱水期）,BnSDP1在低油品系种子中的转录水平高于高油品系,在根、茎、叶、花中均有表达,但在根和叶片中表达量高,在根中尤其显著,且受蔗糖诱导上调表达。     

Potato tuber sugar content during development and storage (10°C): possible predictors of storage potential and the role of sucrose in storage hexose accumulation

Summary Reducing sugar and sucrose contents of potato tubers (Solanum tuberosum L.) from six cultivars were monitored both during the growing season and after a four months storage period at 10°C. Significant correlations were found between tuber sugar contents (sucrose, reducing and total) measured at harvest and the reducing sugar content after storage. Similarly, in five of the six cultivars, the sucrose loss and the corresponding reducing sugar gain during storage were significantly correlated.     

Morphological development and nutritive value of herbage in five temperate grass species during primary growth: analysis of time dynamics

In a 2‐year field experiment, morphological development and measures of the nutritive value of herbage for livestock during primary growth in Meadow foxtail, Tall oatgrass, Cocksfoot, Perennial ryegrass and Yorkshire fog were investigated. All measured variables were affected significantly by both species and sampling date, and their interaction (P < 0·001), in the period of primary growth. Changes with time in mean stage weight for Meadow foxtail and Cocksfoot were different from the other species due to their indeterminate growth habits. Mean stage weight of Tall oatgrass and Yorkshire fog increased more rapidly than that of Perennial ryegrass with time. Changes in mean stage weight with time were described by linear, parabolic and sigmoid relationships. Crude protein (CP) concentration of herbage was higher for Cocksfoot and Meadow foxtail than for Perennial ryegrass. A parabolic relationship of CP concentration with time was typical for all the species. Concentrations of neutral‐detergent fibre (NDF) and acid‐detergent fibre (ADF) in herbage of the species differed most during the mid‐period of primary growth. Their increases with time showed curvilinear (sigmoid and parabolic) relationships. Perennial ryegrass had lower concentrations of both NDF and ADF in herbage than the other species. Differences between the in vitro dry matter (DM) digestibility among the grasses increased in mid‐ and late periods of primary growth. Perennial ryegrass had higher values for in vitro DM digestibility but the difference from other species was small in the early period of primary growth and from cocksfoot in the late period of primary growth. In vitro DM digestibility showed, in most cases, a sigmoid and, in others, a linear decrease with time. Principal component analysis showed that perennial ryegrass and meadow foxtail were the most distinctive of the species in characteristics relating to morphological development and the nutritive value of herbage to livestock.     

Potato mop-top virus (PMTV): distribution in Sweden,development of symptoms during storage and cultivar trials in field and glasshouse

Summary The occorrence of potato mop-top virus (PMTV) was assessed by two different soil sampling methods. PMTV was prevalent in southwestern Sweden but was not detected in the northern part of the country. In storage trials more symptoms developed in tubers kept at 9°C than 4 °C. irrespective of the cultivar. Cutting the tubers enhanced post storage symptom develoment in some cultivars. Of 14 cultivars grown in Sweden, those used for crisp products were more inclined to develop spraing than the others. A comparison of the severity of PMTV symptoms in field trials in Sweden. Denmark and Finland indicated substantial geographic variation in the relative susceptibility of different cultivars suggesting variation among isolates of the virus. From glasshouse experiments it was found that inoculations done at flower bud development or at flowering resulted in most spraing. Cultivar trials in pots produced a low level of spraing symptoms in general.     

Variability and estimating in fruiting of shea tree (Vitellaria paradoxa C.F. Gaertn) associated to climatic conditions in West Africa: implications for sustainable management and development

ABSTRACT

Shea tree (Vitellaria paradoxa) is one of the main native oil plants with high economic value in Africa. Its fruits are used for food, medicinal, cultural and exportation purposes. However, the lack of efficient tools to assess annual fruit production of the species limits the sustainable management of its resources. Therefore, production statistics are useful to organize the activities of the shea sector. This study aimed to (i) assess the interannual variation in fruit production along a climatic gradient in Burkina Faso, (ii) examine fruit production patterns according to climatic zones and tree size, (iii) assess the contribution of high-yielding trees in the annual potential production and (iv) develop allometric equations for estimating fruit yields of the species. The yield of 212 trees distributed across three climatic zones was monitored over four successive years. Within each climatic zone, fruit production was significantly different among years. Fruit production was positively and significantly correlated with tree size. The interannual variation in fruit production at tree individual level was higher in drier climatic conditions. The mean interindividual synchrony was less than 50% in each zone, suggesting a large range in fruit production patterns of the species. Annually, more than 55% of the total fruit production was supported by high-yielding trees. The best regression models for estimating fruit yields had R ² values of 69–73% with prediction errors ranging from ?7% to 4%. Our findings are useful tools for the planning of rational exploitation of shea tree’s resources and its sustainable management.