Analysis of genes related to xylem cell wall development based on transcriptomics in Populus alba ‘Berolinensis’ tension wood

Populus alba‘Berolinensis’is a fast-growing,high-yielding species with strong biotic and abiotic stress resistance,and widely planted for timber,shelter belts and aesthetic purposes.In this study,molecular development is explored and the important genes regulating xylem forma-tion in P.alba‘Berolinensis’under artificial bending treat-ments was identified.Anatomical investigation indicated that tension wood(TW)was characterized by eccentric growth of xylem and was enriched in cellulose;the degree of ligni-fication was lower than for normal wood(NW)and oppo-site wood(OW).RNA-Seq-based transcriptome analysis was performed using developing xylem from three wood types(TW,OW and NW).A large number of differentially expressed genes(DEGs)were screened and 4889 counted.In GO and KEGG enrichment results,genes involved in plant hormone signal transduction,phenylpropanoid biosynthesis,and cell wall and secondary cell wall biogenesis play major roles in xylem development under artificial bending.Eight expansin(PalEXP)genes were identified from the RNA-seq data;four were differentially expressed during tension wood formation.Phylogenetic analysis indicated that PalEXLB1 belongs to the EXPB subfamily and that the other PalEXPs are members of the EXPA subfamily.A transcriptional regulatory network construction showed 10 transcription factors located in the first and second layers upstream of EXP,including WRKY,ERF and bHLH.RT‒qPCR analy-sis in leaves,stems and roots combined with transcriptome analysis suggests that PalEXPA2,PalEXPA4 and PalEXPA15 play significant regulatory roles in cell wall formation during tension wood development.The candidate genes involved in xylem cell wall development during tension wood formation marks an important step toward identifying the molecular regulatory mechanism of xylem development and wood property improvement in P.alba‘Berolinensis’.

Effects of pruning intensity on nonstructural carbohydrates of Populus alba×P.talassica in the arid desert region of Northwest China

Although pruning is important to obtain highquality,large-diameter timber,the effects of pruning on nonstructural carbohydrates(NSC)in aboveground organs of many timber species are not well understood.Three intensities of pruning(none,moderate and severe)were tested on poplars(Populus alba×P.talassica)in the arid desert region of northwest China to compare the concentrations of soluble sugar(SS),starch(ST)and total nonstructural carbohydrate(TNC)in leaves,branches and trunks during the growing season.The concentration of NSC components after different pruning intensities varied similarly in seasonal patterns,increasing slowly at the beginning of the growing season,continuously declining in the middle,then gradually recovering by the end of the growing season.The monthly mean NSC concentration in poplar differed significantly among the three pruning intensities(p<0.05).The SS concentration in pruned trees was higher than in unpruned trees(p<0.05).For moderately pruned trees,the concentrations of ST and TNC in trunks and branches were higher than in unpruned and in severely pruned trees(p<0.05).Compared with no pruning,pruning changed the seasonal variation in NSC concentration.The orders of SS and TNC concentrations in aboveground organs were leaf>branch>trunk,while the order of ST concentration was trunk>leaf>branch,which was related to functional differences of plant organs.The annual average growth in height of unpruned,moderately pruned,and severely pruned poplars was 0.21±0.06,0.45±0.09 and 0.24±0.05 m,respectively,and the annual average growth in DBH were 0.92±0.04,1.27±0.06 and 1.02±0.05 cm,respectively.Our results demonstrate that moderate pruning may effectively increase the annual growth in tree height and DBH while avoiding damage caused by excessive pruning to the tree body.Therefore,moderate pruning may increase the NSC storage and improve the growth of timber species.

An improved draft genome sequence of hybrid Populus alba×Populus glandulosa

Populus alba × P.glandulosa clone 84 K,derived from South Korea,is widely cultivated in China and used as a model in the molecular research of woody plants because of hi gh gene transformation efficiency.Here,we combined63-fold coverage Illumina short reads and 126-fold coverage PacBio long reads to assemble the genome.Due to the hi gh heterozygosity level at 2.1% estimated by k-mer analysis,we exploited TrioCanu for genome assembly.The PacBio clean subreads of P.alba × P.glandulosa were separated into two parts according to the similarities,compared with the parental genomes of P.alba and P.glandulosa.The two parts of the subreads were assembled to two sets of subgenomes comprising subgenome A(405.31 Mb,from P.alba)and subgenome G(376.05 Mb,from P.glandulosa) with the contig N50 size of 5.43 Mb and 2.15 Mb,respectively.A high-quality P.alba × P.glandulosa genome assembly was obtained.The genome size was 781.36 Mb with the contig N50 size of 3.66 Mb and the longest contig was 19.47 Mb.In addition,a total of 176.95 Mb(43.7%),152.37 Mb(40.5%)of repetitive elements were identified and a total of 38,701 and 38,449 protein-coding genes were predicted in subgenomes A and G,respectively.For functional annotation,96.98% of subgenome A and 96.96% of subgenome G genes were annotated with public databases.This de novo assembled genome will facilitate systematic and comprehensive study,such as multi-omics analysis,in the model tree P.alba X P.glandulosa.

Survival and development of immature Harmonia axyridis (Pallas) feeding on Chaitophorus populeti (Panzer) propagated on transgenic Populus alba × P. glandulosa

Laboratory feeding experiments with the poplar aphid, Chaitophorus populeti （Panzer）, feeding on transgenic poplar （P. alba × P. glandulosa） varieties C13-5 and C013-5, were carried out to study the effect of transgenic poplar on the ladybird Harmonia axyridis （Pallas）. The mortality and development time of the immature stages, the eclosion rate and body mass of H. axyridis were measured. The results indicated that C. populeti feeding on different varieties of transgenic plants had no statistically significant ef- fect on the mortality ofH. axyridis larvae. The development time of larval and pupal stages were not significantly different between the two transgenic poplars and a non-transgenic poplar. Furthermore, the body mass and eclosion rate did not show any difference between the H. axyridis feeding on aphids reared on transgenic plants and those from non-transgenic plants. It is suggested that transgenic plants have no deleterious effect on the predatory ladybird.

银中杨、玉簪落叶生物质炭对Pb^(2+)、Cd^(2+)、Cr^(6+)吸附的影响因素

为探讨银中杨、玉簪落叶所制备生物质炭对水体Pb^(2+)、Cd^(2+)和Cr^(6+)吸附规律的差异及影响因素,采用限氧裂解法将银中杨及玉簪落叶制成生物质炭,并以此为吸附载体研究其在不同初始离子质量浓度、pH值、Na+浓度及接触时间等因素影响下对Pb^(2+)、Cd^(2+)和Cr^(6+)的吸附。结果表明:随着初始Pb^(2+)、Cd^(2+)和Cr^(6+)质量浓度的增加(0~800 mg·L^(-1)),落叶生物质炭对相应重金属离子的吸附量也增加。将初始质量浓度设置在0~200 mg·L^(-1),生物质炭对3种金属离子的吸附量由大到小表现为Pb^(2+)、Cd^(2+)、Cr^(6+),然而,将初始离子质量浓度提升至300~800 mg·L^(-1),吸附量由大到小表现为Pb^(2+)、Cr^(6+)、Cd^(2+);溶液pH值由2增至8,可使Pb^(2+)和Cd^(2+)在生物质炭表面的吸附率得到迅速提升,然而,生物质炭对Cr^(6+)的吸附率在整个pH值变化范围则呈渐趋降低的趋势;随着Na+浓度增加(0~0.6 mol·L^(-1)),落叶生物质炭对3种金属离子所表现的吸附规律各不相同,其中,对Pb^(2+)的吸附量先下降而后渐趋升高,对Cd^(2+)的吸附量逐渐下降,而对Cr^(6+)的吸附量则表现为先增加而后下降。Na+离子浓度由0 mol·L^(-1)提升至0.6 mol·L^(-1)可使生物质炭对Pb^(2+)和Cd^(2+)的吸附量分别降低16.8%和97.1%,相反,对Cr^(6+)吸附量却有所促进,使其增加55.6%;生物质炭对初始质量浓度为400 mg·L^(-1)的Pb^(2+)、Cd^(2+)和Cr^(6+)吸附的数量随接触时间延长(0~1 440min)而逐渐增加,相同条件下由大到小表现为Pb^(2+)、Cr^(6+)、Cd^(2+);生物质炭对Pb^(2+)、Cd^(2+)的吸附主要以电性吸附为主,而专性吸附则为生物质炭吸附Cr^(6+)的主要机制。

刺激植物响应蛋白基因Epl1克隆、原核表达及功能初探

【目的】研究刺激植物响应蛋白Epl1对木本植物山新杨的刺激响应功能,为开发新一代提高植物免疫的新型诱导剂奠定基础。【方法】从棘孢木霉ACCC30153菌株中克隆获得一个刺激植物响应蛋白基因Epl1,并进行序列分析和原核表达,将获得的原核重组蛋白rEpl1与山新杨组培苗进行互作,初步探讨rEpl1蛋白对山新杨生长、生理指标、以及生长和防御相关基因转录水平的影响。【结果】刺激植物响应蛋白基因Epl1的cDNA序列全长为417bp,预测蛋白分子量12.6 kDa,属于Cerato-platanin家族;通过构建原核表达载体,成功获得重组蛋白rEpl1;在1μg/mL rEpl1诱导下,山新杨组培苗生长较快、根系旺盛,生物量明显增加,CAT活性在诱导第1 d时为对照的3.51倍,可溶性糖含量和脯氨酸含量在诱导初期急剧积累;山新杨生长素基因LAX2/AUX和水杨酸防御基因JAR2的转录水平分别在蛋白诱导2 d和12 h时达到峰值,分别为对照的873.10和388.02倍。【结论】Epl1基因的克隆、序列分析和原核表达为进一步研究Epl1蛋白诱导植物系统抗病性提供了理论基础;重组蛋白rEpl1与山新杨组培苗互作实验,初步Epl1蛋白能够刺激木本植物在生理及分子水平上的响应,从而促进生长,提高抗性。

银中杨各部位对Cd、Zn、Pb的富集特性

【目的】为探明银中杨各部位对不同重金属的富集特性。【方法】本研究通过用3种不同含量的镉(Cd,0.5、1.5、2.5 mg/kg)、锌(Zn,300、500、700 mg/kg)或铅(Pb,300、500、700 mg/kg)分别处理盆栽一年生银中杨的土壤,分析银中杨根、茎、叶对Cd、Zn和Pb的富集量和富集系数。【结果】银中杨根、茎、叶对Cd、Zn和Pb的富集量均与土壤中相应的重金属含量呈极显著的正相关(P<0.01)。高含量Cd处理下的根、茎、叶对Cd的富集系数均不同程度的低于低含量处理;高含量Zn处理下叶对Zn的富集系数和高含量Pb处理下叶对Pb的富集系数也均不同程度的低于低含量处理,但根、茎对Zn的富集系数对Zn的处理含量不敏感,而根、茎对Pb的富集系数却对Pb的处理含量呈依赖性。银中杨各部位之间对Cd的富集量未表现出任何规律,但对Cd的富集系数,在低含量下一致,在高含量下根显著低于茎和叶(P<0.05)。Zn低含量处理下的叶、根、茎和高含量处理下的根、叶、茎对Zn的富集量和富集系数均依次有不同程度的降低。Pb各含量处理均使根对Pb的富集量和富集系数显著高于茎和叶(P<0.05)。【结论】说明土壤中的Cd、Zn和Pb含量影响银中杨对其的富集,且呈根茎叶特异性。银中杨各部位对Cd的富集情况表现为茎和叶的富集阈限高于根部;对Zn的富集情况在低含量时表现为叶>根>茎,在中、高含量时表现为根>叶>茎;对Pb的富集主要集中在根。

木霉诱导下山新杨PodaPIN9基因的组织表达调控

PIN蛋白具有多个跨膜结构域,影响着高等植物生长素的外向运输和众多生长发育过程。木霉菌是能促进植物生长、提高其对多种病害防御作用的生防因子。研究木霉菌对木本植物山新杨生长素的极性分布的影响有重要意义。克隆了山新杨PodaPIN9基因,对其核酸和蛋白序列进行分析;同时,构建的进化树显示PodaPIN9与6个物种的9条PIN基因具有高度一致性(> 80%)。qRT-PCR分析表明,PodaPIN9在山新杨茎尖、成熟叶和根中均有表达。在根中表达量极低;在茎尖和叶中的表达量极高,分别是根中的503和346倍。该基因受木霉影响在茎尖和叶中的表达量均显著下调;而根中的表达量显著上调,达到对照的32. 01倍。并发现根际接种木霉48h会使杨树茎尖、叶和根中生长素含量降低。说明木霉菌能影响杨树茎尖、叶和根中IAA水平及PodaPIN9的表达量。并且Pearson相关性分析表明在茎尖、叶和根中,PodaPIN9表达量与IAA水平具有不同的相关性。

C/N、pH对银中杨–玉簪落叶及草坪碎屑混料腐解的影响

【目的】以银中杨Populus alba×P.berolinensis–玉簪Hosta plantaginea落叶和草坪碎屑为基础材料,通过对混料C/N、pH的调控,筛选混料腐解的最佳条件。【方法】采用室内培养法对银中杨–玉簪落叶及草坪碎屑混料进行60 d恒温好氧腐解,设置物料C/N为10、15和20以及pH为4、7和10,探讨其对混料腐解过程中水溶性有机碳(WSOC)、可提取腐殖酸碳(HEC)、胡敏酸碳(HAC)、富里酸碳(FAC)及胡敏素碳(Hu C)含量的影响。【结果】将C/N设为10或15时,培养结束后混料HEC及HAC均有所累积,而将混料C/N设为20则更有利于微生物对WSOC、HEC及Hu C的矿化及周转。C/N为15或20时,培养结束后混料腐殖质品质有所改善,而在混料中添加更多氮素使C/N为10时,则在一定程度上阻碍了FAC向HAC的转化。混料C/N为20更有利于其胡敏酸(HA)分子结构的简单化,其次是C/N为10,而C/N为15时更有利于HA分子结构的稳定;混料pH为7有利于微生物对Hu C的矿化,使HEC和HAC含量分别增高30.0%和69.0%。混料pH为10时对HAC/FAC的促进程度最大,使之增加78.3%,pH为4或7时对混料HAC/FAC的促进程度较小。【结论】C/N为15、pH为7时,对银中杨–玉簪落叶及草坪碎屑混料腐解的促进作用最为显著。

银白杨×马氏杨杂交子代及亲本的AFLP分析

为了探讨银白杨、马氏杨及银白杨×马氏杨杂交子代间的遗传多样性参数及个体聚类关系,利用8对引物对银白杨、马氏杨及银白杨×马氏杨杂交子代4个样本进行了AFLP分析。结果表明,8对引物共扩增出780条扩增谱带,其中多态性条带数为715条,多态性比例为91.65%,平均有效等位基因数为1.5017,Nei’s平均遗传多样性指数为0.3065,Shannon平均信息指数为0.4679。不同杂交子代在E-AAC/M-CAG引物对上均有特异性位点,这些特异位点可用于品种鉴定。聚类研究发现,在相似系数为0.76时,试验样品分成2个AFLP群,其中马氏杨为1个AFLP群,银白杨及2个杂交子代为1个AFLP群;当相似系数为0.78时,银白杨和2个杂交子代的AFLP群又划分为2个亚群,其中杂交子代1和杂交子代2聚合为亚群1,银白杨为亚群2。本研究的结果多态性比例很高,对样品的区分率达到100%,为杨树AFLP分析检测提供了指导借鉴价值。

干旱胁迫对银中杨生理生化特性的影响

以银中杨三年生苗木为试验材料,模拟不同程度的土壤干旱环境,研究了土壤干旱对银中杨生理生化特性的影响。结果表明,叶绿素含量在干旱初期降低,持续到干旱14天时,随着干旱程度的加剧,叶绿素含量增加。SOD在轻度干旱胁迫的初期,其活性提高,随着胁迫时间的延长、胁迫强度的加剧,其活性降低,但降低幅度较小;POD活性和可溶性蛋白含量显著降低;类胡萝卜素、脯氨酸含量和电解质含量略微增加,可溶性糖含量显著增加;丙二醛(M DA)含量无明显变化,银中杨叶片的电导率在干旱初期随着土壤含水量的降低而增加,干旱胁迫14天时,差异不显著。

转果聚糖蔗糖转移酶基因银腺杨的获得(英文)

采用农杆菌介导的遗传转化方法,将来自枯草杆菌的果聚糖蔗糖转移酶基因(SacB)导入银腺杨,以提高杨树对水分胁迫的抗性。以来自无菌培养的叶片为外植体,通过大约10 0 0个叶盘与农杆菌LBA4 4 0 4共培养,将植物双元表达载体pKP中SacB基因导入银腺杨基因组,经卡那霉素筛选后,共获得10 2株卡那霉素抗性植株。经PCR特异性扩增和Southern点杂交分析,证明其中97株再生植株基因组DNA中整合了SacB基因。对其中的6 2个无性系进行RT PCR分析,结果表明SacB基因在其中的5 0个无性系中获得表达。温室生长观察表明,转基因无性系外部形态与对照相比没有稳定的显著差异,少数部分转基因无性系的生长明显受到抑制,其他转基因无性系生长正常。这些转基因无性系的获得为培育抗旱转基因杨树奠定了基础。

银中杨光合作用和蒸腾作用对土壤干旱的响应

以银中杨3年生苗木为试验材料，通过盆栽试验，研究土壤干旱程度、干旱持续时间和逐渐干旱对银中杨叶片气体交换参数的日变化和光响应过程的影响。结果表明：银中杨具有较强的抗旱性；适度干旱有利于银中杨光合作用；逐渐干旱过程中，土壤含水量在40．1％-20．2％范围内，光合速率随着含水量的减少而降低，但降低幅度较小，表观量子效率基本不变；土壤含水量为15．5％时的光合能力则大幅度下降；银中杨的光饱和点在750～1300μmol·m^-2·s^-1之间，随着土壤含水量的下降，光饱和点降低；光合速率和腾速率与气孔导度的正相关性较强，与胞间CO2则有负相关的趋势；在逐渐干旱过程中，水分利用率随着含水量的下降而上升，但长时间的严重干旱胁迫导致水分利用率降低；在相同含水量条件下，逐渐干旱对各光合参数的影响均小于梯度干旱，这说明银中杨光合作用能忍受暂时的干旱，重度水分胁迫持续时间较长时，光合速率和蒸腾速率大幅度降低，这是银中杨对干旱的一种适应方式。

银腺杨与毛新杨正反交三倍体选育

采用花粉染色体人工加倍获得的银腺杨 (Populusalba×P .glandulosa)、毛新杨 (P .tomentosa×P .bolleana)未减数 2n花粉 ,施加一定剂量的辐射处理后杂交 ,两杂交组合共获得 16株三倍体 .其中银腺杨×毛新杨组合以1470rad处理最佳 ,三倍体得率 3 .8% ;而毛新杨×银腺杨 (16 80rad)则获得了高达 12 .9%的三倍体 .三倍体整体生长水平优势明显 ,单株生长更为突出 ,2年生实生苗的苗高和地径最高 4.9m和 5 .8cm ,分别超出群体平均值的5 8%和 176 % ;并且正反交三倍体的遗传效应差异明显 .利用未减数 2n花粉 ,经花粉辐射处理杂交选育杂种三倍体 ,将是今后白杨乃至杨属植物育种的最有效途径之一 .

转基因银腺杂种杨对土壤微生物的影响

In this study,genetic stability of foreign genes was checked by using PCR technology. A total of 4 lines of soil samples from transgenic and non-transgenic control Populus alba×P.glandulosa and the soil samples from the stands were collected in two successive years,and the bacteria,fungi and antinomyces were isolated by using flat dilution method to determine their quantities. The results demonstrated that target genes were stable present in the genome of transgenic poplar. We found that three kinds of soil microorganism appeared different trends within different months and different years. The ANOVA and multiple comparison analyses revealed that there was no significant difference in quantity of the soil microorganisms among most poplar lines at the same time point. Meanwhile,significant differences of the soil microorganism community were found among several transgenic lines,and between a few transgenic and non-transgenic lines,but they did not show any relation to the type and number of foreign genes. Our preliminary results indicated that the 2-year-old transgenic poplar growing in field had no significant effects on soil microorganisms.

外源SacB基因在银腺杂种杨基因组中的表达及抗旱性分析

利用转基因技术培育抗旱杨树新品种是改善干旱地区生态环境的有效途径之一.该研究对已整合外源SacB基因的银腺杂种杨株系进行半定量RT-PCR和温室水分胁迫试验.研究结果表明,在所测定的7个转基因株系中,外源SacB基因均成功转录mRNA;转基因株系叶片中积累了SacB基因表达产物(果聚糖);在干旱胁迫条件下,转基因株系的生长量、生物量和叶片含水量明显高于对照.相关分析表明转基因株系生长量、生物量和叶片含水量与果聚糖积累量呈极显著正相关,说明SacB基因的导入提高了转基因杨树对水分胁迫的抗性.

土壤水分供应梯度对银中杨扦插生根及苗木生长的影响

对不同土壤供水条件下银中杨插穗萌发、生根及生长进行研究 ,结果表明 :土壤水分状况对银中杨穗条萌发影响不显著 ,但影响生根率和苗木生长。干旱胁迫条件下 ,插穗生根率不如中等供水处理和淹水处理。中等供水处理扦插苗在水分利用效率、苗木生长及生物量方面 ,显著高于淹水和干旱胁迫处理。综合插穗生根及扦插苗生长 ,银中杨扦插育苗以中等供水处理 (土壤含水量 31.7% )

一株能拮抗3种土传病害病原真菌的长枝木霉

为了丰富生防木霉菌(Trichodermaspp.)种资源,在园林植物八宝景天(Sedum spectabile)根际土壤中分离得到一株优势木霉。根据该菌株菌落的形态特征、形态学显微观察、菌株rDNA-ITS序列分析以及邻接法同源性比对等结果判定木霉菌株种类。采用平板对峙法将该菌株与3种土传病害病原菌[(Sclerotinia sclerotiorum)、立枯丝核菌(Rhizoctonia solani)和细链格孢菌(Alternaria alternata)]进行对峙培养。并用该木霉诱导山新杨(Populus davidiana×P.albavar.Pyramidalis)组培苗,观察离体叶片抵抗细链格孢菌侵染的能力。结果表明,此木霉为长枝木霉(Trichoderma longibrachiatum),命名为Tl-70。该菌株对3种土传病害病原真菌均有较明显的拮抗作用。对核盘菌的抑菌率最高为77.71%,显著高于对细链格孢菌和立枯丝核菌的抑制率(P<0.05);对立枯丝核菌的抑菌率为58.56%,显著高于对细链格孢菌的抑制率(P<0.05);对细链格孢菌的抑菌率最低,为53.32%。并且,该木霉能提高山新杨组培苗叶片拮抗细链格孢菌侵染的能力。说明此菌株是具有潜力的生防菌株。

转vgb基因银腺杂种杨的耐涝性

对导入透明颤菌血红蛋白(vitreoscilla hemoglobin,VHb)基因vgb银腺杂种杨进行Southern印记杂交和RT-PCR检测,证实vgb基因已整合到杨树基因组中,并在转录水平上得到表达。在温室对4个转基因株系及对照盆栽扦插苗进行模拟持续淹水胁迫试验,研究淹水条件下植株的生长及生理性状。结果表明,随着淹水时间的延长苗木逐渐死亡,对照植株在淹水17天后死亡率达66.67%,而4个转基因株系死亡率均低于50.00%,其中T81最低,只有25.00%;同时4个转基因株系在淹水胁迫时能够保持较高的叶绿素含量,并且膜质过氧化物MDA含量均低于对照植株;涝渍胁迫下转基因株系生长受抑制程度低于对照植株。说明在淹水胁迫下vgb基因的表达增强了银腺杂种杨叶绿素含量的积累,进而促进了转基因植株的生长,提高了转基因株系对淹水的忍受能力。

多效唑对银中杨、白榆和白桦抗旱性的影响

在干旱胁迫条件下，植物会产生一系列的生理变化，从而影响其生长发育（杨建昌等，2004；时连辉等，2005）。从研究的对象来看，主要集中在应用高效、低毒植物生长调节剂提高农作物或1年生植物的抗旱性上（曹翠玲等，2004；周凤珏等，2004），而在林木，尤其是城市绿化树种上的应用研究较少（彭立新等，2004；温国胜等，2004）。多效唑是一种高效低毒的植物生长延缓剂。国内外对多效唑在农作物、果树、蔬菜、花卉上的作用和应用已进行了广泛研究（Kraus et al．，1995；汪志辉等，2003；周凤珏等，2004；阮龙等，2005；