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.

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。

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。

不同水溶肥对马铃薯生长发育及产量的影响

为使马铃薯(Solanum tuberosum L.)生产提质增效,研究3种水溶肥对马铃薯生长、产量、品质以及商品率的影响。以马铃薯希森6号为试验材料,共设4个处理,分别为F0(不施水溶肥,对照)、F1(腐殖酸水溶肥)、F2(微生物水溶肥)、F3(中药水溶肥),测定不同水溶肥处理下马铃薯形态、地上部分干物质累积量、产量及品质等相关指标。结果表明,追施水溶肥能提高马铃薯的株高、茎粗、叶绿素SPAD及地上部分干物质累积量,并能增加产量与提高品质。以微生物水溶肥的表现最突出,较不施水溶肥株高增加5.23 cm,单株块茎数提高19.92%,干物质累积量增加35.47%,商品率增加10.95%,产量提高27.13%、淀粉含量增加11.87%,维生素C含量增加16.08%。由此可知,微生物水溶肥不仅可以促进马铃薯生长,增加产量,而且能够显著提高马铃薯品质,可作为当地灌溉追施的高效水溶肥。

环洞庭湖地区绿芦笋套作马铃薯免耕栽培技术

绿芦笋因其丰富的营养以及脆爽的口感,深受消费者青睐,栽种收益多年且单年收益高,是较好的富民产业。绿芦笋冬季剪枝时间与马铃薯生产时间重叠,适合开展绿芦笋与马铃薯套作模式。主要从绿芦笋的品种选择、田间准备、中期管理、病虫害防治、嫩笋采后处理分级等方面以及马铃薯品种选择、田间准备、中期管理、病虫害防治、收获与收益等方面总结了绿芦笋套作马铃薯免耕栽培技术要点,并就该套作模式的优点进行了分析,以期为广大种植者提供参考。

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。

马铃薯DHN基因家族的鉴定与分析

脱水素(dehydrin,DHN)作为一种高亲水性蛋白,在植物响应各种生物、非生物胁迫过程中发挥重要作用。本研究在马铃薯基因组中鉴定出7个StDHNs基因,不均匀地分布在3条染色体上,编码氨基酸数目为80~243 aa,内含子数目均为1;StDHNs蛋白分子量在8.54~25.12 kDa之间,等电点为5.24~7.38,共3个保守motifs,保守基序及进化关系分析表明StDHN家族成员在进化上具有高度的保守性。转录表达分析结果显示,StDHN5、StDHN6、StDHN7在特定逆境胁迫下显著高表达。诱导表达模式分析表明,StDHN5、StDHN6受到盐和ABA胁迫的显著诱导,StDHN7响应干旱胁迫。本研究结果可为深入研究马铃薯DHN基因家族的功能及其在逆境胁迫抗性中的应用提供科学依据。

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。

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。

磷转运蛋白StPHO1.2提高马铃薯耐热性

植物通过磷转运蛋白吸收和转运磷元素,充足的磷元素能够提高作物产量、品质和抗逆性。高温是影响马铃薯生长发育的重要环境因子,严重时会造成减产甚至绝收。本研究利用农杆菌介导,在马铃薯(Solanum tuberosum L.)中过表达磷转运蛋白基因StPHO1.2,比较转基因株系和野生型株系在高温(35℃)和常温(22℃±1℃)环境下的生长状况。结果表明,过表达StPHO1.2能够提高马铃薯耐热性,促进其生长,且磷元素浓度越高,抗性越强,长势越好。本氏烟草中的亚细胞定位结果显示,StPHO1.2在细胞膜上表达,因此,选择膜系统文库质粒筛选StPHO1.2的互作蛋白。通过酵母双杂交试验和BiFC试验,本研究证明磷转运蛋白StPHO1.2与钙离子转运相关蛋白(StCAX1)和光系统Ⅱ蛋白亚基(StPsbR)均有相互作用。综上所述,过表达StPHO1.2可能通过影响光合作用和信号转导,从而提高马铃薯耐热性,促进马铃薯生长。这些结果为深入理解磷转运蛋白的功能提供了理论依据和参考,对马铃薯新品种的选育具有促进作用。

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。

施钾对马铃薯苗期抗旱性的影响

为研究不同施钾水平对马铃薯苗期抗旱性的影响,本研究以马铃薯品种大西洋为材料,对苗期不同干旱胁迫的大西洋增施不同量钾肥,测定其生长指标和生理指标,并探究在不同条件下植株的抗旱性差异。结果表明:干旱胁迫会显著抑制马铃薯株高、茎粗的生长,同时显著降低叶片叶绿素含量、叶片类胡萝卜素含量、光合生理指标及根系活力,会促使马铃薯叶片中丙二醛含量、脯氨酸含量、抗氧化物酶活性增加,进而影响马铃薯块茎的产量及品质。增施一定量的钾肥可有效缓解干旱对上述指标的影响,均在K2(180 kg/hm^(2))水平下效果最佳。在K2施钾水平下,T0、T1和T23种干旱胁迫条件下的马铃薯块茎重量较未施钾肥的分别增加了23.3%、27.8%和36.2%,块茎淀粉含量分别增加了25.2%、25.47%和33.67%,维生素C含量分别增加了34.83%、36.91%和43.05%。综上,施钾量为180 kg/hm 2对马铃薯幼苗抗旱性和生长发育的促进效果最佳。

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.

马铃薯后代材料晚疫病的抗性鉴定

为筛选出农艺性状优良且抗病性较好的后代材料,本试验对60份马铃薯“青薯9号”与“大西洋”杂交后代材料的离体叶片进行4种不同菌株的接种,分析4种菌株接种后的病斑面积及抗性等级,并利用LAMP技术对接种不同天数的离体叶片进行扩增检测。结果表明在60份后代材料中,高抗有6份,抗性有11份,中抗有17份,感病有24份,高感有2份。表现抗病的后代材料一共有34份,均具有广谱性,其中18份后代材料对4种菌株均产生抗病,16份后代材料对3种或2种菌株产生抗病,对4种菌株表现抗性的后代材料的广谱性比对3种或2种菌株表现抗性的后代材料高。利用LAMP技术检测接种1~5 d内及5 d后离体叶片的晚疫病菌,其扩增检测结果一致,均呈蓝色,为阳性。综上,本研究筛选的马铃薯后代材料可为抗病育种提供品种参考,LAMP技术能够对马铃薯晚疫病达到早期鉴定及预警的目的,为马铃薯晚疫病早期防治提供技术支持。

微量植物组织汁液直接RT-LAMP可视化检测马铃薯卷叶病毒方法的构建

为建立快速的马铃薯卷叶病毒检测方法以应用于脱病毒后大规模样本的检测,构建利用微量植物组织汁液直接逆转录环介导恒温扩增(RT-LAMP)可视化检测马铃薯卷叶病毒的方法,即以无菌大头针的针尖刺马铃薯植株茎尖时所粘附的汁液直接作为RNA模板,采用Bacillus stearothermophilus(Bst) DNA聚合酶进行RT-LAMP扩增,扩增产物通过肉眼观察有无白色沉淀来诊断马铃薯卷叶病毒.该方法快速、操作简单、成本低,在大规模组织培养无病毒种薯生产中具有很大的应用空间.

青海地区马铃薯主要病毒检测与分析

病毒病是影响马铃薯产量和种薯质量的重要因素之一。为了解青海省马铃薯主要病毒种类,本研究于2019—2020年从青海省湟中区、大通回族土族自治县、湟源县、互助土族自治县和乐都区生产田中采集疑似病株叶片245份,从不同级别种薯上采集种薯201份。应用酶联免疫吸附法(DAS-ELISA)对马铃薯PVY、PVX、PLRV、PVA、PVM、PVS和AMV等7种主要病毒进行筛查。结果表明:在105份样品中检测到病毒,其中PVY病毒的检出率最高,为28.98%;PVS的检出率次之,为21.22%;PLRV、PVX、PVM和PVA的检出率相对较低,分别为5.31%、4.90%、4.49%和2.86%;未检测到AMV。2种病毒复合侵染较普遍,主要类型为PVY+PVS,侵染率达到10.20%;未发现3种及以上病毒复合侵染现象。在不同县区马铃薯生产田中病毒种类和带毒率差异明显。不同级别种薯的病毒检测结果符合马铃薯种薯质量标准。

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.