The Impact of High Temperature during Growing Season on Potato Cultivars with Different Response to Environmental Stresses

Potato crop is the fourth main food crops in the world after maize, rice and wheat. It is characterized by specific temperature requirements and develops best at about 20°C. Forecasts of global warming prompt us to study the tolerance of potato genotypes to heat during the growing season. The aim of this work was to assess the response of chosen potato cultivars to high temperature during the different stages of plant growth under conditions of good soil moisture and drought. The impact of high temperature 32°C/25°C on potato plants was determined in pot experiment in three growth stages. A main measure of tolerance of the potato cultivars to high temperature during the growing season was an evaluation of the yield in relation to the Control combination. Here we demonstrated that tested potato cultivar’s response to high temperature during the growing season is dependent on the growth stage. The earlier it occurs, the more negative its impact on the growth and yield of potatoes is.

Isolation of a Plasmalemma Aquaporin Encoding Gene StPIP1 from Solanum tuberosum L. and Its Expression in Transgenic Tobacco

Aquaporin （AQP） belongs to a highly conserved group of membrane proteins considered as major intrinsic proteins, which facilitate water transport across biological membranes. The discovery of AQPs in plants has resulted in a paradigm shift in the understanding of plant-water relations, however, the potential relationship between the role of aquaporins in regulating plant water balance and drought tolerance still remains elusive. In this study, the gene encoding potato AQP cDNA, StPIP1 （GenBank accession no. DQ999080）, was cloned from the leaf of potato cultivar Gannongshu 2 by reverse transcription-PCR （RT-PCR）. Sequence alignment was made by BLASTn in GenBank, the phylogenetic analysis was conducted using PHYLIPWY, the 3D structure was predicted in Swiss-Model server. Subcellular localization of StPIP1 was performed by constructing CaMV35S-StPIP1-GFP and rd29A-StPIP1-GFP fusion proteins and transient expression in onion epidermis. To understand StPIP1 physiological functions in potato under various stress conditions, the StPIP1 gene in a reverse orientation was transformed into tobacco driven by the Cauliflower mosaic virus （CMV） 35S promoter. The expression levels of transgenic and wild-type plants were assessed under various abiotic stress conditions using semi-quantitative RT-PCR, and the morphological and physiological responses of transgenic plants to different stress conditions were investigated. The expression of StPIP1 mRNA decreased in transgenic plants under non-stress and stress conditions, however, the reduction was more severer under drought stress. In both non-stress and stress conditions, StPIP1 was expressed predominantly in root. The morphological and physiological investigation showed no significant differences in growth rate, germination rate, and root fresh weight （FW） between transgenic and wild-type plants when grown under favorable conditions. In contrast, under drought stress, the reduction in StPIPI expression leads to a delay in seed germination and seedling growth, accelerated seedling wilt, and leaf morphological abnormity. Under ＂enough＂ water conditions （i.e., water culture）, the aerial parts of anti-sense plants showed no differences. However, for the aerial parts to accumulate the same amount of biomass, transgenic plants needed about 3 times more abundant root system to transport water for plant growth than wild-type plants. Morphological investigation showed that the reduction in StPIP1 expression increased the root system in transgenic plants under drought stress. As a result, the increase of root mass might compensate the reduced cellular water permeability in order to ensure a sufficient water supply for the plant. Results demonstrated that StPIP1 plays an important role for water transportation in potato, especially under drought stress conditions. The reduced expression of StPIP1 decreases the cellular water transport and influences the expression of endogenous AQPs genes and thereby, has impacts on seed germination, seedling growth, and stress responses of potato to drought conditions.

马铃薯PYL5基因对非生物胁迫的响应分析及其启动子的活性鉴定

【目的】脱落酸(ABA)作为一种“应激激素”在植物生长发育和响应干旱、盐等非生物胁迫过程中发挥重要作用,PYR/PYL/RCARs(以下简称“PYL”)作为ABA受体在多种植物中也被广泛研究。基于对马铃薯StPYL5基因生物信息学及表达模式分析,并通过对其启动子活性鉴定,为进一步揭示马铃薯StPYL5功能及抗逆育种提供依据。【方法】根据转录组数据克隆得到StPYL5基因,通过DNAMAN、MEGA等软件分析了StPYL5的分子特征;通过qPCR检测了StPYL5基因的组织特异性及其对非生物胁迫的响应;利用PlantCARE网站对StPYL5基因启动子进行了分析,并通过瞬时转化烟草对其活性进行了鉴定。【结果】StPYL5基因全长534 bp,共编码177个氨基酸,蛋白质分子量20.19 ku,理论等电点(pI)为5.97。系统进化分析显示,StPYL5与SpPYL9-like亲缘关系较近。组织特异性分析结果显示,青薯9号(QS9)中StPYL5在根和叶中表达量较高,其次分别是茎和花,在块茎中表达量较低。不同胁迫下StPYL5表达量分析表明,青薯9号(QS9)中StPYL5在干旱、低温、盐和ABA胁迫下的表达量先升高后降低,且StPYL5的表达受Me JA和SA的诱导。此外,笔者成功克隆得到2 000 bp的StPYL5基因启动子。在烟草中的瞬时转化后的组织化学染色结果表明,StPYL5基因启动子具有成功启动下游GUS报告基因表达的启动子活性。【结论】全面分析了StPYL5基因的分子特征及其在多种非生物胁迫下的表达谱,并成功克隆到具有活性的pStPYL5启动子,该结果为深入研究StPYL5基因的功能以及马铃薯抗逆育种提供依据。

马铃薯块茎RNA提取及RT-PCR法克隆酸性转化酶基因

马铃薯块茎由于富含多糖和酚类物质造成RNA提取难度大。通过在SDS-phenol提取缓冲液中加入2%PVP及10mmol/L半胱氨酸以除去大量的酚类物质,利用改进后的SDS-酚法提取的马铃薯块茎总RNA电泳结果表明,总RNA质量高于未经改进的方法。并进行RT-PCR克隆与马铃薯块茎低温糖化反应过程密切相关的块茎酸性转化酶(acidinvertase)基因,经核酸序列对比分析,与公布的已知序列具有98.96%的同源性,为利用反义技术抑制酸性转化酶活性进行马铃薯块茎抗低温糖化研究作好了准备工作。

Effects of different planting systems on yield of potato crop in Kaghan Valley: A mountainous region of Pakistan

Conversion of potato from conventional methods to wide bed planting systems may increase water and nitrogen use efficiency in commercial potato production system by reducing the amount of irrigation water and water applied nitrogen fertilizer bypassing the potato root zone. Potato (Solanum tuberosum L) cv. Desiree was tested against different planting system for yield and yield components at Kaghan, a high mountainous Himalayan region ofPakistan. The experiment was carried out at Himalayan Agricultural Research Station (HARS), Kaghan during the summer season of 2005. The results showed that maximum tuber growth (88.7%), number of stems per plant (3.5), plant expansion (45.5 cm), average number of tubers per plant (10.1) and yield per hectare (12.4 t/ha) were significantly different and higher when potatoes were planted on wide bed and covered with soil from one side. Tallest plants (53.4 cm) were observed when potatoes were sown on the ridges. Maximum number of green potatoes (12.5) and injured potatoes (5.3%) were observed when the tubers were planted following local farmers’ method. Keeping in view the soil type, land slopping, we recommend sowing potatoes on relatively plain wide beds and covering it with soil from one side, for potato cultivation in the area.

Transcriptome analysis reveals effects of red and blue lightemitting diodes(LEDs)on the growth,chlorophyll fluorescence and endogenous plant hormones of potato(Solanum tuberosum L.)plantlets cultured in vitro

Red and blue light illumination has been reported to significantly affect plantlet growth.Potato is an important food and feed crop in the world and potato plantlet cultured in vitro plays an important role in potato production.However,few studies have documented the effects of red and blue light on the growth of potato plantlets revealed at the transcriptome level.The objective of this study was to determine the growth and physiological responses of potato plantlets cultured in vitro under monochromatic red(RR),monochromatic blue(BB)as well as combined red and blue(RB)LEDs using the RNA-Seq technique.In total,3150 and 814 differentially expressed genes(DEGs)were detected in potato plantlets under RR and BB,respectively,compared to RB(used as control).Compared to the control,the DEGs enriched in"photosynthesis"and"photosynthesis-antenna proteins"metabolic pathways were up-regulated and down-regulated by BB and RR,respectively,which might be responsible for the increases and decreases of maximum quantum yield(F_(v)/F_(m)),photochemical quantum yield(φ_(PSII)),photochemical quenching(q_(P))and electron transfer rate(ETR)in BB and RR,respectively.Potato plantlets exhibited dwarfed stems and extended leaves under BB,whereas elongated stems and small leaves were induced under RR.These dramatically altered plantlet phenotypes were associated with variable levels of endogenous plant hormones gibberellin(GAs),indoleacetic acid(IAA)and cytokinins(CKs),as assessed in stems and leaves of potato plantlets.In addition,monochromatic red and blue LEDs trigged the opposite expression profiles of DEGs identified in the"plant hormone signal transduction"metabolic pathway,which were closely related to the endogenous plant hormone levels in potato plantlets.Our results provide insights into the responses of potato plantlets cultured in vitro to red and blue LEDs at the transcriptomic level and may contribute to improvements in the micro-propagation of potato plantlets cultured in vitro from the light spectrum aspect.

组织培养条件下马铃薯茎段对NaCl基础抗性的研究

本文以马铃薯(Solanum tuberosum L.)东农303和Favorita两个基因型茎段为材料,研究了在组织培养条件下NaCl胁迫对马铃薯不同基因型茎段芽与根的形成以及幼苗生长的影响。结果表明,东农303和Favorit两个基因型对NaCl的基础抗性存在差别,依基因型不同NaCl筛选浓度分别为75mmol/L和100mmol/L。本研究明确了马铃薯两个基因型茎段在组织培养条件下对NaCl的基础抗性,为进一步筛选马铃薯耐盐植株奠定了基础。

Class I patatin基因5′侧翼区驱动的GUS基因在马铃薯块茎中专一性表达

将1.4kb Class I patatin基因的5′侧翼区与GUS基因融合,构建了双元表达载体pPATIs(含patatin部分信号顺序)和pPATI(不含patatin部分信号顺序)。pPATI通过基因枪介导在块茎切片中获得了瞬间表达。以上建构物通过农杆菌介导转入了马铃薯品种Desiree。X-Gluc染色(PATIs不能染色)及PCR结果证实已获得转基因植株。利用离体块茎诱导系统,GUS的表达进一步用荧光进行定量检测,结果显示,PATI-GUS的转基因植株中GUS比活性均以块茎明显高于茎段,达10-20倍。蔗糖浓度的升高,PATI-GUS植株中的GUS比活性无明显变化,与前人报道有不同。此外,光照促进PATI-GUS的表达。

高温和利巴韦林对马铃薯的生长及PLRV、PYV的影响

为了了解高温和利巴韦林处理对马铃薯(Solanum tuberosum L.)卷叶病毒(Potato leave-roll virus,PLRV)和马铃薯Y病毒(Potato Y virus,PYV)脱毒的影响,以宝塔百利、红王和"803"为材料,采用高温和化学处理结合茎尖培养技术对上述马铃薯进行处理,采用RT-PCR方法检测病毒基因。结果表明,长时间的高温处理抑制马铃薯组培苗的生长,"803"的存活率最高为80.1%,宝塔百利的存活率最低为41.1%;利巴韦林对马铃薯生长的抑制作用较明显,其浓度超过75 mg/L时生根效果差,到125 mg/L时对马铃薯生长抑制效果最明显;RT-PCR结果显示,高温处理结合利巴韦林的处理上均未检测到病毒。

异属砧木嫁接对茄子和番茄生长、产量及品质的影响

嫁接是克服茄果类蔬菜连作障碍的重要手段,砧木是影响嫁接蔬菜生长、产量和品质的关键因素之一。试验分别采用番茄属(Lycopersicon Mill.)和茄属(Solanum L.)砧木对番茄(Lycopersicon esculentum Mill.)及茄子(Solanum melongena L.)进行了嫁接,研究嫁接后对番茄和茄子生长、产量及品质的影响。结果表明,异属砧木和接穗嫁接亲和力相对较低。番茄属砧木嫁接番茄对其生长无影响,但能显著增加茄子的株高、茎粗和叶片数;茄属砧木显著降低了番茄的生长势,减少了叶片数,显著增加了茄子的株高和叶片数。番茄属砧木嫁接番茄后可显著降低番茄的糖酸比,降低茄子的可溶性糖含量;茄属砧木显著降低了番茄的可溶性糖含量,增加了可滴定酸含量,降低了糖酸比,增加了茄子维生素C和粗纤维含量。番茄属砧木显著提高了番茄和茄子的产量,茄属砧木显著降低了番茄的产量,显著增加了茄子的产量。因此,在生产上建议,茄果蔬菜嫁接采用同属植株为砧木比较合适。

龙葵正丁醇提取物对Lewis肺癌移植瘤生长的影响

目的:观察龙葵正丁醇提取物对Lewis肺癌移植瘤生长的抑制作用。方法:建立Lewis肺癌模型,将40只C57BL/6J荷瘤小鼠随机分成荷瘤空白对照组、环磷酰胺组、龙葵正丁醇提取物高、中、低剂量组5组,每组8只。给药15 d后处死小鼠,剥离肿瘤组织,称重,计算肿瘤生长抑制率;解剖其胸腺、脾脏组织,称重,计算胸腺和脾脏指数。结果:龙葵正丁醇提取物高、中、低剂量对Lewis肺癌移植瘤的抑瘤率分别为32.14%、13.93%和15.36%。龙葵正丁醇提取物高剂量组胸腺指数增长率为134.31%,龙葵正丁醇提取物中剂量组脾指数增长率为128.73%。结论:龙葵正丁醇提取物对Lewis肺癌移植瘤生长具有一定程度的抑制作用,并能提高荷瘤小鼠胸腺和脾脏指数。

Impact of Organic Soil Amendments on the Physical Characteristics and Yield Components of Potato （Solanum tuberosurn L.） in the Highlands of Cameroon

A field experiment was conducted in the Western Highlands of Cameroon to study the effects of Calliandra calothyrsus, sterilized compost, non-sterilized compost and mineral fertilizers （NPK 11：11：22） on physical characteristics, yield components and late blight disease severity of potato （Solanum tuberosum）. A two factorial treatment combination made up of fertilization schemes and sanitary measures were laid out in a randomized complete block design （RCBD） with four replicates and 10 treatments. Data collected were subjected to a multivariate ANOVA and means separated with the Dunnett t-test with Calliandra calothyrsus considered as the main treatment. Mineral fertilizers and Calliandra calothyrsus significantly augmented stem diameter, plant height and plant vigor, total and marketable yields, despite the fact that late blight severity was high in the two treatments. The correlation matrix showed that total yield had significant and positive correlation with stem diameter （r = 0.74）, plant height （r = 0.61）, plant vigor （r = 0.61） and marketable yield （r = 0.99） and negative correlation （r = -37） with late blight severity. The increase observed is attributed to the nutrient contents of the treatments applied. Leafy prunings of C. calothyrsus increase soil organic matter which probably improves moisture retention and biological activity. The highest late blight severity recorded between the 45 and 60 coincides with active development of plant canopy which influences disease severity and create a canopy microclimate conducive to disease development. This study suggests that Calliandra calothyrsus has the potential of improving potato production provided appropriate fungicide is used.

Planting Adaptability of Four Kinds of Common Vegetabless in Shanghai

[Objectives]To explore the planting adaptability of vegetables in Shanghai.[Methods]In this paper,cowpea(Vigna unguiculate(L.)Walp.),cucumber(Cucumis sativus L.),eggplant(Solanum melongena L.)and potato(Solanum tuberosum L.)were selected as experimental materials and planted in the open air.The growth status,the occurrence of diseases and insect pests,and the taste evaluation of these four kinds of common vegetables were mainly studied.[Results]The results showed that the four kinds of common vegetables in Shanghai had strong growth,strong adaptability,less pests and diseases,and good taste.[Conclusions]The cowpea,cucumber,eggplant,and potato are suitable for planting in Shanghai.

马铃薯PPR蛋白家族基因SoDIPPR的克隆及其在干旱条件下的表达特征分析

【目的】PPR(pentatricope ptide repeat)蛋白家族在植物的生长发育、细胞器形成、细胞质雄性不育的育性恢复、RNA的编辑加工、细胞核与细胞器之间信号传递、逆境防御等方面具有重要作用。研究旨在了解PPR蛋白家族SoDIPPR基因在马铃薯中的表达情况及其在干旱胁迫下的作用【。方法】以抗旱马铃薯二倍体品系H145为材料,利用cDNA-AFLP技术寻找与抗旱性密切相关的基因cDNA片段,利用RACE技术克隆其基因全长cDNA,并利用半定量RT-PCR和Northern杂交法研究其在干旱条件下的表达情况。【结果】从抗旱马铃薯二倍体品系H145中克隆了一个836bp全长cDNA序列,命名为SoDIPPR,该序列开放阅读框长为588bp,编码195个氨基酸序列。序列分析表明,SoDIPPR蛋白存在PPR结构域、激酶结合区、和C端SMR(small MutS related protein)区域。SoDIPPR蛋白序列与GenBank数据库中的其它PPR蛋白进行同源序列比对,构建系统进化树,发现SoDIPPR与其它14个高等植物的PPR蛋白同源性为57%～82%,与苜蓿DNA错配修复蛋白MuTs2、水稻盐诱导蛋白(Q9LS25)和叶绿体RNA结合蛋白(Q75IP8)的同源性达69%～82%。半定量RT-PCR和Northern杂交结果表明,SoDIPPR基因在干旱胁迫下叶片和根系里的表达量明显增加,说明SoDIPPR基因在马铃薯抗旱中起一定作用。在持续干旱条件下,SoDIPPR基因在抗旱品系H145与干旱敏感品系H214中表达模式不同。【结论】马铃薯SoDIPPR基因编码的蛋白与其它植物PPR家族蛋白同源性较高,SoDIPPR基因参与马铃薯对干旱胁迫的应答反应,可能对抵抗干旱胁迫有一定作用。

转Aclnv反义基因马铃薯品系的生理特性及POD同工酶变化分析

对转Aclnv反义基因马铃薯品系的相对电导率、丙二醛含量(MDA)和超氧化物歧化酶(SOD)活性进行检测.结果表明:转基因"大西洋"品系的相对电导率显著低于受体品种,且C2代的MDA含量显著低于受体水平;转基因"台湾红皮"品系相对电导率和MDA含量与受体品种无显著差异;Aclnv反义基因的导入对"大西洋"和"台湾红皮"两个马铃薯品系的SOD含量影响不大.对转Aclnv反义基因马铃薯品系过氧化物酶同工酶谱带分析表明,转Aclnv反义基因马铃薯品系与受体亲本存在明显的变异.

The Key Factors Affecting Tuber Development of Potato in vitro and the Relation with Protein Fractions

According to previous analysis, some properties bounding up with tuber yield were investigated. The results showed that tuber average weight, plastid Mg2 + -ATPase activity, plastid Ca2 + -ATPase activity, mitochondria Mg2 + -ATPase activity, total soluble protein content, tuber average diameter, and Q-enzyme activity were important factors determining the tuber yield. The linear regression equation was:Y = 0.5211 + 0.0595X(1)+0.8389X(2) +0.0882X(3) -0.0073X(4) +0.1449X(5) +0.3510X(6) +0.0031X(7) -0.00003X(8) + 0.3412X(9) + 0.0127X(10) + 0.2904X(ll) + 0.0570X(12) + 0.0159X(13) + 0.3585X(14) + 0.0134X(15) - 0.1012X(16). At the same time, the relation between several important properties and soluble protein fractions were analyzed.

Genetic Variance Estimates in Heterogeneous Potato Populations Propagated from True Potato Seed(TPS)

Twenty nine cross populations developed according to line × tester mating design along with their twelve parents raised from true potato seeds, were used to study variability and correlation for seven characters. The range of variation for all the characters were found to be wide and pronounced which indicated that characters were quantitative in nature and were under polygenic control. For all the characters, greater portion of the total δ2p was mostly contributed by the δ2g. Among the seven characters, tuber weight per plant showed the highest δ2g, PCV, GCV, H2b, GA and expected GA%. On the other hand, dry matter contents (%) showed the lowest H2b. Phenotypic and genotypic correlation coefficients of plant height with no. of tubers per plant and tuber weight per plant; no. of branches per plant with no. of tubers per plant and tuber weight per plant were found to be significant and positive.

茄子及其近缘野生种AFLP分析体系的建立

本文以若干茄子自交系及其近缘野生种为材料,从DNA提取、酶切连接、预扩和选扩等几方面进行选择优化,建立起高效稳定的AFLP分析体系;从64对AFLP引物组合筛选出多态性好的引物组合38对,栽培种茄子和野生种托鲁巴姆间多态性高达68.5%。本实验室正以本AFLP分析体系开展茄子资源遗传多样性、青枯病抗性基因分子标记等遗传分析研究工作。

马铃薯StCML基因家族鉴定及表达分析

类钙调蛋白(calmodulin-like protein,CML)是植物中一种重要的Ca2+结合蛋白,在植物生长发育和胁迫响应过程中起着重要的作用。该研究通过生物信息学方法在马铃薯基因组中鉴定了StCML基因家族成员,并对它们的表达模式及胁迫响应进行了分析,为深入解析马铃薯StCML基因家族成员在生长发育和胁迫响应中的作用机制奠定理论基础。结果显示:(1)在马铃薯基因组中共鉴定到80个StCML基因,它们均具有EF-hand结构;根据系统进化树拓扑结构可分为5个亚家族,在1~5亚家族中分别含有18、12、14、12、和24个基因,大部分基因具有较为保守的基因结构和基序。(2)RNA-Seq数据分析发现,StCML基因主要在马铃薯的花、叶柄、芽、雄蕊、匍匐茎和块茎中有特异表达,并且主要对盐、热、干旱和赤霉素处理有响应。(3)qRT-PCR分析发现,在低温胁迫下StCML13、StCML21和StCML53表达上调;在高温胁迫下StCML11、StCML21和StCML39表达上调;盐胁迫下StCML21和StCML60表达上调;青枯菌处理下StCML53表达上调,StCML8、StCML 13、StCML 21和StCML 60表达下调。研究表明,StCML基因对多种胁迫均有响应。

Chromosome Number Manipulation as Part of Potato Pre-breeding Programs

The cultivated potato(Solanum tuberosum L.) is a tetraploid(2n=4x=48) and can be improved with the incorporation of desirable traits from other Solanum species.Often the transfer of these traits is hindered by complex genetics and breeding barriers within potato.Parthenogenesis and microsporogenesis are used in chromosome number manipulation allowing breeders to reduce the potato's chromosome number to dihaploid(2n=2x=24)[diploid] or monohaploid(2n=x=12) from which a predictable transfer of traits can be made,in accordance with the endosperm balance number theory(EBN).Furthermore,the reproductive processes of first division restitution(FDR) and second division restitution(SDR) are utilized in order to increase the chromosome number for incorporation into the cultivated potato.