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.

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。

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.

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.

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.

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.

The Role of Biodiversity in Food Security and Nutrition: A Potato Cultivar Case Study

Biodiversity is considered a critical measure of the agricultural health of the world. Not only does increased biodiversity contribute to nutrient production and consumption, but it acts as a safeguard against food shortages due to pests and diseases by spreading the risk. Biodiversity can improve dietary diversity in such a way to ultimately contribute to improved food and nutrition security. As a result biodiversity is often highlighted in global discussions related to food and nutrition security. Potatoes (Solanum tuberosum L.) are highly biodiverse food crops, with more than 4000 different cultivars grown globally. The crop is sometimes considered as part of the vegetable component of food baskets, but it is mainly added to meals as a starch because potatoes have a high starch content and are thus high in energy. However, significant differences in the nutritional content (including micronutrient composition) of different potato cultivars have been reported in many countries. It is therefore proposed that specific cultivars could potentially contribute more to critical nutrients required in the diet of countries at risk of malnutrition. The nutritional profile (macronutrients and minerals) of 11 potato cultivars cultivated in sub-Saharan Africa was determined and the contribution which these different potatoes could make to the diet, and consequently to food and nutrition security, is discussed. The results showed that significant differences are found in the nutritional content between the different cultivars. Significant differences (p < 0.001) were found in macronutrients such as protein and fat as well as micronutrients such as copper, phosphorus and potassium. These results indicate the potentially beneficial role which a biodiverse range of crops such as potatoes, could play in the food and nutrition security of developing countries.

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.

Effects of Agrobacterial<i>rol</i>-Genes on the Thermodynamic and Structural Features of Starches Extracted from Potato Microtubers

Wild-type potato (Solanum tuberosum L.) plants and their transformants harboring agrobacterial rolB or rolC genes under control of the patatin class I promoter were cultured in vitro. These plants were used as a source of single-node stem cuttings. The structure of native starch in tubers formed on cuttings was determined using methods of X-ray scattering and differential scanning microcalorimetry (DSC). It was found that in starch from tubers of rolB plants the melting temperature of crystalline lamella was lower and their thickness was less than that in wild-type potato. In tubers of rolC plants starch differed from starch in wild-type plants by a higher melting temperature, reduced melting enthalpy, and a greater thickness of crystalline lamellae. The melting of starch from tubers of rolC plants proceeded as the melting of two independent crystalline structures with melting temperatures of 338.0°K and 342.8°K. Overall data show that starches of different structure can be obtained by using transgenic approach.

Analysis of Diversity among Potato Accessions Grown in Eritrea Using Single Linkage Clustering

Potato is an important cash and food crop, a source of income to many small scale farmers in Eritrea. In the long tradition of potato cultivation farmers have adopted a number of landraces. However, very little is known about their genetic diversity which limits further breeding programs to improve productivity. The objective of the study was to characterize potato accessions grown in Eritrea. A total of 21 accessions collected from farmers and NARI were used in this study for characterization employing 33 phenotypic descriptors. Single linkage (nearest neighbor) clustering method using UPGMA employing Euclidean test was used to yield a dendrogram. The clustering generated three main clusters with further sub clusters. The clustering helped to identify accessions that shared the same characteristics and/or that were closely related and vice versa. It was noted that there was no distinct relationship between the clusters groups and geographic sources of accession. Yet, accessions sharing the same name may not necessarily share the same genetic content. Mean deviation of variables for each group from the total mean indicated that Group III followed by group I showed negative deviation in most of the recorded yield related traits. Group I was particularly affected by the presence of two inferior accessions in the group. On the other hand, Group II accessions showed relatively maximum deviation from the total mean and could be recommended for use as parent for any breeding program in Eritrea.

Preservation of Disease-free Shoot Tips of Potato Germplasm through in vitro System

The present investigation was undertaken to find out the best medium composition for medium-term preservation of disease free potato shoot tips in in vitro system.Thirteen potato genotypes and fourteen treatments were taken under consideration for the present experiment.Among fourteen treatments,mannitol and sorbitol containing media proved to be the best for medium-term preservation of potato shoot tips.High concentration of mannitol delayed root formation.

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.

Rooting Potential of Mini-Cuttings for the Production of Potato Plantlets

The production of mini-tubers under soilless cultivation system increased the availability of pathogen-free seed potatoes. The objective of this study was to evaluate the rooting capability of mini-cuttings of potato clones as a function of the irrigation solution and the physiological age of the mother plant, under soilless cultivation system. Potato mini-cuttings were collected from young and mature plants of the cultivars Asterix and Macaca and the advanced clone SMINIA 793101-3. The young plants were newly acclimatized individuals and the mature plants were at the stage of producing mini-tubers. In the first experiment, mini-cuttings originating from summer pruning of mature plants were grown under closed soilless cultivation system irrigated with a nu- trient solution developed for the production of potato mini-tubers and a solution comprised of tap water only. The experiment was conducted in a 3 × 2 factorial arrangement (clones and irrigation solutions) in the complete random design with four replicates of 15 mini-cuttings. In the second experiment, the young and mature mini-cuttings were established under a closed soilless cultivation system irrigated with nutrient solution. The experiment was conducted in a 3 × 2 factorial arrangement (clones and physiological age of the mother plant) in the complete random design with four replicates of 15 mini-cuttings. In both experiments, the percentage of rooting and survival of mini-cuttings, number of roots and length of the longest root were assessed at 21 days of cultivation. Potato mini-cuttings can be rooted under closed soilless cultivation system irrigated with nutrient solution or tap water. Mini-cuttings of young plants have higher rooting capability compared to the ones taken from mature plants. The loss of rooting capability depends upon the potato cultivar.

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.

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。

华北寒旱区马铃薯的氮效率评价及筛选

为筛选氮高效马铃薯品种,提高冀西北地区氮肥利用效率,本试验以15个马铃薯品种为供试材料,设置大田条件下不施氮与施氮两种处理,从马铃薯块茎产量、氮浓度、氮吸收量、氮生物学效率、氮肥农学效率、氮肥偏生产力、氮肥表观利用率等方面进行氮效率评价;并采用隶属函数对各马铃薯品种进行综合氮效率评价及筛选。结果表明,品种与施氮水平均会影响马铃薯产量,在不施氮处理下大西洋的产量最高,干重为10645.70 kg/667 m^(2);施氮处理下丽薯产量的增幅最大,为23.38%。15个马铃薯品种在施氮肥产量、氮吸收量、氮肥农学效率、氮肥偏生产力及氮肥表观利用率等氮效率性状评价下存在宽幅变异,变异系数为7%~64%。施氮与不施氮条件下,马铃薯氮浓度与产量均无显著相关关系,氮肥表观利用率与氮肥农学效率呈显著正相关。隶属函数分析表明,马铃薯品种大西洋、冀张薯8号、北方002、丽薯的综合氮效率较高,可利用于生产或氮高效种质资源。

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。

马铃薯WOX基因家族的鉴定及在离体再生和非生物胁迫中的表达分析

【目的】WUSCHE-相关同源盒(WUSCHEL-related homeobox,WOX)基因家族是植物特有的转录因子家族,在植物生长发育、干细胞分化调控、逆境胁迫响应等过程中扮演重要角色。开展马铃薯WOX基因家族鉴定与功能研究,将为马铃薯遗传改良提供优良基因资源与理论依据。【方法】基于拟南芥、番茄、烟草和水稻WOX蛋白序列,利用HMMER 3.0和BLASTP鉴定马铃薯WOX基因家族成员,使用MCScanX软件分析WOX基因家族成员在马铃薯种内及种间的共线性,并采用邻接法构建系统发育进化树。利用ExPASy、GSDS等软件分析马铃薯WOX基因家族成员理化性质、基因结构、蛋白motif、启动子区域转录因子结合位点。基于PGSC数据库中马铃薯转录组数据,分析StWOXs在不同组织和非生物胁迫下的表达模式;以可能参与离体再生过程的StWOX5作为候选基因,利用实时荧光定量PCR技术分析该基因在具有不同离体再生能力的4个马铃薯品种(系)再生过程中的表达情况。【结果】鉴定得到11个马铃薯WOX基因家族成员,分布在5条染色体上,分为WUS、中间和古老共3个进化分支,不同分支中StWOXs基因结构、蛋白motif组成、转录因子结合位点类型和数量存在不同程度的差异。表达模式结果表明,马铃薯WOX基因家族成员参与离体再生和非生物胁迫响应,StWOX4/5/11/13在愈伤组织中特异表达,StWUS、StWOX1/3c/4/5/13在甘露醇处理下上调表达,StWOX3a/11在NaCl处理下上调表达,StWOX4/5/13在热胁迫中上调表达,且StWOX5相对表达量与愈伤组织分化率呈正相关。【结论】不同马铃薯WOX基因具有潜在的功能多样性,StWOX5具有促进马铃薯离体再生分化的作用,参与了非生物胁迫反应。