Genetic Variability, Heritability and Correlation of Some Morphological and Yield Components Traits in Potato (Solanum tuberosum L.) Collections

The study was conducted with the main objective to evaluate the genetic variability, heritability, and clustering pattern exploration of morphological and yield related traits in potato (Solanum tuberosum L.) collections in the bimodal rainfall agroecological zone of Cameroon using a Randomized Complete Block Design (RCBD) with three replications. The data obtained on morphological and yield traits were subjected to analysis of variance (ANOVA). The results showed that the viability rate of the collections varied from 77.78% to 96.55% respectively for the Maffo and Desiree collections, while the greatest number of tubers per plant varied from 4 to 18 respectively for Synergie and Desiree. The emergence rate varies from 60% to 1.66% respectively for Maffo et Doza collections. However, Desiree presents the highest TL (96.55) while Maffo shows the lowest value (77.78%). The yield per hectare varied from 1.14 to 9.3 t/h for Maffo and Doza respectively. For all the characteristics observed, Phenotypic Coefficients of Variation (PCV) were higher than Genotypic Coefficients of Variation (GCV) suggesting the role of environment in the expression of traits under observation. The highest GCV and PCV 47.55 and 58.94 respectively were observed for Diameter at the collar (DC). Most of the traits showed high GAM (>20%)) except Average Tuber Length (ATL) with a moderate value (19.8). In terms of vegetative development, the Desiree variety showed the highest performance. Based on the growth and yield results, Doza seems to be the most recommendable crop in the study 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.

Environmental, morphological and physiological factors analyzes for optimization of potato (<i>Solanum tuberosum</i>L.) microtuber <i>in vitro</i>germination

The microtuber is considered one of the most effective means of spreading basic materials, as well as transporting and preserving potato germplasm varieties. To define the optimal conditions for the potato microtuber in vitro germination of Aida, Atlas and Odessa varieties, the effects of temperature, physiological age and grade (size) were evaluated. The study conducted at three different temperature levels has demonstrated that the most favorable temperature for microtuber germination at a higher and faster germination rate was 25℃, regardless of the variety. In addition, microtubers of large caliber, greater than 4 mm, germinate more quickly, with a higher germination rate, than smaller size ones (less than 4 mm) for all genotypes. For Atlas, Aida and Odessa varieties, a germination rate equal to 86.66%, 70% and 70% respectively, was obtained for microtubers with a caliber superior to 4 mm. Physiological age influences microtuber germination. The mean length of sprouts, reached after a 7 week incubation period, was more marked at “multiple sprout” and “branched sprout” stages than at a “monosprout” stage. The average length was 2.35 cm, 2.48 cm and 1.5 cm, respectively. Thus, it is necessary to plant microtubers at a “multiple sprout” stage to optimize their yield in plants and minitubers.

Residual Effects of Sucrose and Hormonal Treatments of the Tuberization Medium on <i>in Vitro</i>Germination of Potato (<i>Solanum tuberosum</i>L.) Microtubers

The residual effects of sucrose concentrations (80 or 100 g·L-1) and hormonal treatments (BAP + Kinetin or Coumarin) of tuberization medium on in vitro microtubers germination of three potato varieties (Solanum tuberosum L.) so called Aida, Atlas and Odessa, are described. After 3 weeks of incubation at 28℃ ± 1℃, 70% of Aida microtubers variety, previously formed in the MT2 medium [MS/2 + 80 g·L-1 Sucrose], germinated. The best germination rate for varieties Atlas (100%) and Odessa (66.66%) was obtained on microtubers previously formed in the medium MT2 [MS/2 + 100 g·L-1 Sucrose]. The addition of hormones in the tuberization medium allowed optimizing the microtubers germination of the Aida variety unlike the other varieties. Indeed, for the Aida variety, the combination M5 [Kin 2.5 mg·L-1 + Coum 0.025 mg·L-1 + Sucrose 80 g·L-1] increased the germination rate from 70% up to 93.33%. The best germination rate (90%), noticed with microtubers of Atlas variety, initially formed in M2 medium [Kin 1 mg·L-1 + BAP 1 mg·L-1 + Sucrose 100 g·L-1], was lower than that one (100%) obtained on medium without hormones. For Odessa variety, the maximum germination rate (53.33%) of microtubers, from the medium M4 [Kin 2.5 mg·L-1 + BAP 1 mg·L-1 + Sucrose 100 g·L-1], was also lower than that one (66.66%) observed in the medium without hormones. Aida and Atlas varieties thus offer a better germination rate than Odessa after their cold storage.

Preliminary Findings on Yield Response of Potatoes （Solanum tuberosum） to Umostart Super Zn （11.46,0 ＋ 2 Zn ＋ Humates）

Field trials to determine the yield response of potatoes to Umostart Super Zn （USZ） fertilizer （11.46.0 ＋ 2 Zn ＋ Humates） were carried out in 2004 using the cultivar Tigoni. During the long rains season, the treatments included USZ at 15 kg ha^-1 at planting; 30 kg ha^-1 split into 15 kg ha^-1 at planting and 15 kg ha^-1 during final earthing-up; 30 kg ha^-1 at planting; and 45 kg ha^-1 at planting. Treatments during the short rains included USZ at 30 kg ha^-1 split into 15 kg ha^-1 at planting and 15 kg ha^-1 during final earthing-up; 30 kg ha^-1 at planting; 40 kg ha^-1 at planting; 50 kg ha^-1 at planting; 50 kg ha^-1 split into 25 kg ha^-1 at planting and 25 kg ha^-1 during final earthing-up; and 60 kg ha^-1 at planting. USZ was contrasted with Diammonium phosphate （DAP） fertilizer （18：46：0） at 300 kg ha^-1 and 500 kg ha^-1 applied at planting and a control in which no fertilizer was applied. The DAP 300 kg ha^-1 treatment was however, not included during the long rains season. The highest tuber yields among the USZ treatments were obtained in plots treated with 45 kg ha^-1 （27.11 t ha^-1） during the long rains and 60 kg ha^-1 （55.17 t ha^-1） during the short rains season. Application of USZ resulted in a yield increase of at least 3.90 and 19.06 t ha^-1 during the long and short rains seasons respectively.

Genetic Diversity and Yield Stability of Potato(Solanum tuberosum L.)

The present study was under taken to investigate the genetic distance and the performance of thirty one potato genotypes in diverse environments.Multivariate and canonical analysis were used to study genetic diversity among the genotypes and stability parameters were estimated according to Eberhart and Russell,Parkinge and Jinks and Freeman and Perkins models for plant height,branch number per plant,tuber number and tuber weight.Thirty one genotypes were grouped in to six clusters with the help of Mahalonobis distance and canonical analysis.The stability analysis revealed that yield was greatly influenced by the environments.Most of the studied genotypes were sensitive for yield with the environment.For over-all performance 'Hera','Chamak','Patnai','Lala pakri','TPS-7','TPS-364','Somerset' and 'Superior' were the best genotypes for tuber yield and breeding material for potato breeding.

Influence of the Size of Potato Microtubers (<i>Solanum tuberosum</i>L.) on the Yield of Plants under Semi-Axenic Conditions

The potato (Solanum tuberosum L.) is a vegetable that ranks fifth in the world for human consumption. Its importance is growing more and more in the Senegalese diet. However, the potato production in Senegal does not meet the needs of the market, which maintains dependence on the outside for the supply of quality seeds. In addition, these imported seeds do not often have phytosanitary qualities required for local production in the Sahelian zone. The in vitro production of microtubers, used as seed, has been shown to be one of the most efficient means for propagation of basic material. To overcome the constraints linked to the supply and availability of potato seeds, with high germination capacity, the impact of the microtuber size on the yield of the plants under semi-controlled conditions was examined. The pre-germinated microtubers were produced in vitro from vitroplants of 3 different varieties (Aida, Atlas, Odessa) adapted to the edaphic-climatic conditions of Senegal. The effects of the seed sizes of microtubers, greater than 4 mm, sown under semi-controlled conditions, on the yield of the plants, result in an increase in the ratio, in the vegetative development of the plants, but also in the number and size of the minitubers harvested. The yield of the plants also depends on the variety. It can therefore be envisaged to produce local potato seeds from microtubers and minitubers.

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.

Molecular Cloning and Expression Analysis of a FT Homologous Gene from Solanum tuberosum

A homologue of flowering locus T gene, designated StFT, was isolated from Solanum tuberosum by reverse transcriptasepolymerase chain reaction （accession no. GU223211）. The DNA sequence of StFT was 1 626 bp long and contained four exons and three introns. The open reading frame of the gene was 522 bp long and encoded a putative protein of 173 amino acids with a molecular weight of 19.75 kD and a theoretical pI of 7.76. StFT protein had a conserved PBP domain and a higher degree of identity with FT homologous members from other species. Analysis on the mRNA levels of StFT showed that it was highly expressed in leaves, apical buds, flowers, and swelling stolons. Further analysis indicated that its expression was regulated by CONSTANS gene in StCOL-antisense transgenic potato plants.

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.

Growth of Micropropagated Solanum tuberosum L.Plantlets under Artificial Solar Spectrum and Different Mono-and Polychromatic LED Lights

In agriculture,LED light sources have increasingly replaced the standard luminescent lamps and have acquired an important role in plant micropropagation.We studied the effect of different light sources such as narrow-band LEDs(bright blue,blue,green,yellow,deep red,and red)and wide-band LEDs(cold white,white,warm white,full spectrum,and an artificial solar spectrum sun box constructed by us)on development of potato plantlets in vitro.White luminescent lamps were used as a control.The light intensity of 49μmol·m^(-2)·s^(-1)was provided in all light treatments.We showed that the long-wave narrow-band light treatments were inapplicable for potato micropropagation,because plantlets were weak with small leaves,inhibited roots,and significantly elongated stems.Blue lights provided growth of shortened plantlets with large leaves,well-growing roots,and abundant green mass.The chlorophyll content was lower under blue and bright blue light and was at the same level in the remained treatments.Significant differences in the stomatal apparatus development were observed depending on the light source.These differences were not always reflected in the plantlet phenotype:e.g.,plantlets under blue and bright blue lights showed no differences in any characteristics except stomatal density and size of stomatal guard cells.We found no significant effect of blue light portion in the white lights and full spectrum on plantlet growth.An artificial solar spectrum sun box was the most suitable for potato micropropagation,because it supported the development of plantlets with good fitness,uniform internodes length,abundant roots and green mass accumulation.

马铃薯野生种烯酰水合酶超家族基因ScDHNS的克隆与功能分析

【目的】1,4-二氢氧-2-石脑-CoA合成酶(1,4-dihydroxy-2-naphthoyl-CoA synthase,DHNS)基因是茄科植物糖苷生物碱合成代谢的潜在重要基因,开展马铃薯DHNS基因功能研究与验证,为低糖苷生物碱马铃薯品种(系)的选育提供基因和材料来源。【方法】利用RACE方法克隆得到马铃薯野生种恰柯薯(Solanum chacoense)ScDHNS基因,对其进行生物信息学分析和亚细胞定位,通过构建过表达载体pBWA(V)HS-DHNS转化马铃薯栽培种进行功能验证。【结果】ScDHNS cDNA序列开放阅读框1023 bp,编码340个氨基酸,分子量为37.34 kD,等电点pI为8.592,具有典型的ECH保守结构域,属于烯酰水合酶超家族成员,在二穗短柄草(Brachypodium distachyon)、蒺藜苜蓿(Medicago truncatula)等植物基因组中都有其同源基因,且存在基因扩张和收缩事件。过表达ScDHNS基因后发现转化株ScDHNS和SGT1基因表达量显著上调,且表达量显著高于马铃薯WT植株。且对应转化植株的总糖苷生物碱含量显著高于马铃薯WT植株,最高可达到364.3 mg/kg,是对照的2.4倍。亚细胞定位结果显示ScDHNS定位于过氧化物酶体。【结论】马铃薯ScDHNS基因可能参与调控糖苷生物碱合成关键基因SGT1的表达,通过β-氧化途径和甲羟戊酸通路协同影响糖苷生物碱的合成,该基因与糖苷生物碱在亚细胞水平上的区室化有重要关系,对于培育低糖苷生物碱的马铃薯品种(系)具有重要的应用价值。

‘怀玉山’高山马铃薯叶绿体基因组特征及密码子使用偏好性分析

【目的】分析‘怀玉山’高山马铃薯Solanum tuberosum var.cormosus‘Huaiyushan’叶绿体基因组特征及密码子使用偏好性,为开展‘怀玉山’高山马铃薯叶绿体基因组密码子优化、叶绿体基因组改造,探索物种进化和增加外源基因表达等研究提供参考依据和理论基础。【方法】采用高通量测序技术对‘怀玉山’高山马铃薯叶绿体基因组进行测序,并利用生物信息学分析软件对组装和注释后的叶绿体基因组进行结构、基因组成及密码子偏好性分析。【结果】‘怀玉山’高山马铃薯叶绿体基因组大小为155296 bp,为经典的4段式结构。大单拷贝区(LSC)、小单拷贝区(SSC)和反向重复区(IR)长度分别为85737、18373、25593 bp,总鸟嘌呤和胞嘧啶所占的比例(GC比例)为37.88%,共注释出133个基因,包含87个编码区(CDS)基因、37个tRNA基因、8个rRNA基因和1个假基因。‘怀玉山’高山马铃薯叶绿体基因组中共检测到38个简单重复序列位点(SSR位点,36个单碱基重复和2个双碱基重复)和32个长重复序列(16个正向重复和16个回文重复)。‘怀玉山’高山马铃薯叶绿体基因组核苷酸多样性为0-0.13927,高变区主要分布在大单拷贝区和小单拷贝区,大单拷贝区trnL-UAA-trnF-GAA、cemA、rps12-exon1-clpP1、clpP1基因变异率最高,小单拷贝区rpl32-trnL-UAG、ycf1基因变异率最高。‘怀玉山’高山马铃薯叶绿体基因组87个CDS基因的平均有效密码子数(ENC)为47.29,ENC>45的基因有60个,密码子偏性较弱。‘怀玉山’高山马铃薯叶绿体基因组密码子偏好以A、U结尾,使用偏性很大程度上受自然选择的影响,而受突变压力的影响小。CGU、AAA、CUU、GUU、GGA、GUA、GGU、UCA、GCU、CCU为‘怀玉山’高山马铃薯叶绿体基因组的10个最优密码子。【结论】‘怀玉山’高山马铃薯与马铃薯栽培种S.tuberosum‘Desiree’亲缘关系较近。

StGID1基因调控马铃薯植株高度、直径和叶绿素含量

为了研究赤霉素不敏感矮化基因(gibberellin insensitive dwarf,GID1)在马铃薯生长中的作用,在马铃薯中克隆该基因并转化植株记录相关表达差异。以马铃薯‘川芋21’组培苗为材料,通过马铃薯基因库设计引物克隆到了一个马铃薯赤霉素不敏感矮化基因StGID1。通过转化‘鄂马铃薯3号’,得到过表达转基因植株和RNAi植株并记录表达差异。进化分析显示,StGID1蛋白氨基酸序列与番茄SlGID1b-1基因的氨基酸序列同源性最高。过量表达和干扰抑制表达的转基因植株对比显示,在组培苗中,干扰抑制表达的植株平均茎直径为过量表达植株的2倍。在成熟植株中,抑制表达植株株高仅为过表达植株株高的62.8%,显著低于空白对照和过表达植株。在赤霉素处理时,过表达转基因植株、干扰抑制表达植株及对照均表现出随赤霉素浓度升高而茎高增高,茎直径减小。本研究证明StGID1是GID1基因家族在马铃薯中的同源基因,为马铃薯赤霉素调控路径的一部分。

暹罗芽胞杆菌JZ1-4-10对马铃薯干腐病的抑菌作用及机理

为了得到对马铃薯干腐病具有良好抑菌效果的生防细菌,本研究从4株芽胞杆菌中筛选出暹罗芽胞杆菌JZ1-4-10,并通过马铃薯活体试验探究菌株JZ1-4-10防治马铃薯干腐病及其相关防御酶活性的影响。结果表明,4株芽胞杆菌中菌株JZ1-4-10的抑菌活性最高,对茄病镰刀菌的抑菌率达59.15%,发酵液的抑菌率达61.25%;此外,菌株JZ1-4-10处理有效提高了马铃薯SOD和POD活性,降低PPO酶活性和MDA含量,保持了马铃薯贮藏期间的品质,且对马铃薯干腐病的预防效果优于治疗、钝化的防治效果,达85.17%;同时,该菌株菌悬液及挥发性有机物均有较高的抑菌活性,并具有较强的生物膜形成能力。因此,暹罗芽胞杆菌JZ1-4-10对马铃薯干腐病有着巨大的防治潜力,可进一步开发成生防菌剂。

80份国际马铃薯中心引进的马铃薯种质资源抗旱性鉴定及评价指标的筛选

【目的】中国马铃薯种质资源相对缺乏,遗传背景单一,亲缘关系较近是制约我国马铃薯种质创新与优良新品种选育的重要原因。此外,干旱是马铃薯(Solanum tuberosum L.)生产的主要影响因素之一。因此,筛选出优良且抗旱的种质资源不仅能够有效应对干旱带来的马铃薯生产挑战,也将为解决中国马铃薯遗传背景狭窄和种质资源短缺提供重要的解决途径。【方法】以从国际马铃薯中心(CIP)表型观测评价的TON panel约300份高世代品系种质资源中筛选出80份表现良好的四倍体种质资源为试验材料,于甘肃省定西市马铃薯研究所进行田间试验,对正常灌溉(CK)和干旱处理(D)下的15个主要农艺性状、产量性状和品质性状进行测定。利用相关性分析、主成分分析、隶属函数分析、聚类分析和逐步回归等分析方法对80份马铃薯种质资源的抗旱性进行综合评价。【结果】通过计算各单项指标的抗旱系数(PI)和变异系数(CV)筛选到12个代表性指标进行后续分析。通过主成分分析,将12个单项指标的抗旱系数转换为6个相互独立的综合指标(累计贡献率达80.45%),根据综合评价值(D值)和聚类分析,可将80份马铃薯种质资源材料分为3类:B20、B40、B55等9个品系属于I类高度抗旱类型;B32、B77、B28等62个品系属于Ⅱ类中度抗旱类型;B73、B42、B19等9个品系属于Ⅲ类低抗类型。进一步利用逐步回归分析建立了抗旱性评价回归模型Y=-0.138+0.198PH+0.080RL+0.109LAI+0.144RFW+0.162TDW+0.079RSC(R2=0.917,P=0),估计精度均在86.6%以上,表明该模型可用于马铃薯抗旱种质资源筛选。【结论】B20、B40、B55等9个品系为高度抗旱品系,可作为优良的杂交亲本选育马铃薯抗旱品种。株高、总根长、叶面积指数、根总鲜重、单株薯块干重、还原糖含量可作为快速筛选抗旱马铃薯种质资源的预测指标,使马铃薯抗旱性评价鉴定工作更为便捷。

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。

马铃薯种质资源抗黑痣病鉴定及SSR聚类分析

马铃薯黑痣病已成为影响马铃薯产量和商品价值的重要土传病害。为丰富马铃薯抗黑痣病种质资源,提高抗黑痣病育种工作效率,研究通过54份马铃薯种质资源地下茎与匍匐茎黑痣病抗性评价,共筛选出7份中抗资源,分别为克17-33-270、龙18-124-8、龙18-7-100、ND11-7-22、克18-134-5、克17-115-1、克17-6-9。研究利用8对SSR适宜引物对7份抗性材料DNA进行PCR扩增,获得SSR多态性条带42条,多态性比率89.36%,遗传相似性系数在0.444 4~0.833 3之间,以相似性系数0.62为基准,可将7份抗性材料分为3类,其中克18-134-5和克17-115-1为一类;龙18-124-8和龙18-7-100为一类;克17-33-270、ND11-7-22和克17-6-9为一类。

茉莉酸调控马铃薯离体块茎发育的主要代谢物变化

【目的】茉莉酸(jasmonic acid,JA)是马铃薯块茎发育过程中重要调控激素之一,研究JA调控块茎发育机理将为块茎产量及品质性状形成提供重要理论基础。【方法】采用马铃薯离体匍匐茎作为供试材料,外源添加JA(0、0.5、5和50μmol·L^(-1))处理,分析JA诱导离体块茎表型形态、组织显微结构、碳水化合物积累及蛋白质组变化。【结果】随JA处理浓度升高,单个匍匐茎形成块茎数目、块茎直径及干鲜重、环髓区细胞面积、淀粉及可溶性糖含量在0.5和5μmol·L^(-1)JA处理后逐渐增加(P<0.05),而50μmol·L^(-1)JA处理后块茎直径及干鲜重、淀粉含量显著减少(P<0.05);脂氧合酶活性随JA浓度升高逐渐降低(P<0.05)。采用双向凝胶电泳(2-DE)和MALDI-TOF/TOF-MS质谱技术鉴定到JA调控块茎发育密切相关的35个差异表达蛋白质(P<0.05且差异表达倍数≥2.5倍),主要涉及生物能量与代谢(28.6%)、细胞防御(28.6%)、蛋白质生物合成与贮藏(11.4%)、信号转导(8.6%)、转录(8.6%)、未知功能(8.6%)和混杂(5.6%)。通过聚类分析将这些蛋白质差异表达模式聚为三类:第一类包括17个蛋白质,在0.5μmol·L^(-1)JA处理后下调表达,而在5μmol·L^(-1)JA处理后上调表达,主要参与生物能量与代谢、蛋白质生物合成与贮藏、信号转导、转录;第二类包括10个蛋白质,随JA浓度升高上调表达,主要参与细胞防御、生物能量与代谢、转录;第三类包括8个蛋白质,JA处理后均下调表达,主要参与生物能量与代谢、细胞防御、蛋白质生物合成与贮藏。【结论】低浓度JA(0.5和5μmol·L^(-1))主要通过诱导块茎环髓区细胞膨大、细胞内蔗糖及多糖积累、细胞防御能力来促进块茎形态建成,而高浓度JA(50μmol·L^(-1))则表现为抑制作用。

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。