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1.
The transport of silicon is an integral part of the synthesis of the silicified cell wall of diatoms, yet knowledge of the number, features, and regulation of silicon transporters is lacking. We report the isolation and sequence determination of five silicon transporter (SIT) genes from Cylindrotheca fusiformis, and examine their expression patterns during cell wall synthesis. The encoded SIT amino acid sequences are highly conserved in their putative transmembrane domains. Nine conserved cysteines in this domain may account for the sensitivity of silicon uptake to sulfhydryl blocking agents. A less conserved C-terminal domain is predicted to form coiled-coil structures, suggesting that the SITs interact with other proteins. We show that SIT gene expression is induced just prior to, and during, cell wall synthesis. The genes are expressed at very different levels, and SIT1 is expressed in a different pattern from SIT 2–5. Hybridization experiments show that multiple SIT gene copies are present in all diatom species tested. From the data we infer that individual transporters play specific roles in silicon uptake, and propose that the cell regulates uptake by controlling the amount or location of each. The identification of all SIT genes in C. fusiformis will enhance our understanding of the mechanism and control of silicon transport in diatoms. 相似文献
2.
M. Torki P. Mandaron F. Thomas F. Quigley R. Mache D. Falconet 《Molecular & general genetics : MGG》1999,261(6):948-952
By systematic sequencing of a flower bud cDNA library from Arabidopsis thaliana, we have identified four cDNAs encoding polygalacturonase. The corresponding genes, together with seven other A. thaliana genes present in the databases, form a small gene family. Sequence comparisons of the deduced polypeptides within the gene
family or with other plant polygalacturonases allow classification of the genes into different clades. Five polygalacturonases,
including all those isolated from the flower buds, are closely related to the enzyme in pollen. Of the six remaining polygalacturonases,
three are more closely related to the abscission-specific type of enzyme and two others to the fruit polygalacturonase. The
last one is more distantly related to the others and might correspond to a new type of polygalacturonase. Expression of the
different genes was analysed on Northern blots and by a PCR-based strategy. Results indicate that if, as expected, the cDNAs
isolated from the flower bud library are strongly expressed in pollen, other genes are expressed at a low level in young developing
tissues, such as in seedlings and roots, suggesting that they could be implicated in the cell wall modifications observed
during cell elongation and/or expansion which occur in these tissues.
Received: 7 December 1998 / Accepted: 1 April 1999 相似文献
3.
Pectin demethylesterification appears to be catalysed by a number of pectin methylesterase (PME) isoenzymes in higher plant species. In order to better define the biological role of these isoenzymes in plant cell growth and differentiation, we undertook molecular studies on the PME-encoding genes in Arabidopsis thaliana. In this paper, we report the characterization of AtPME3, a new PME-related gene of 4 kb in length that we have mapped on Chromosome III. AtPME3 encodes a putative mature PME-related isoenzyme of 34 kDa with a basic isoelectric point. Since the extent of the gene family encoding PME in higher plant species is still unknown, we resorted to the use of degenerate primers designed from several well-known consensus regions to identify new PME-related genes in the genome of Arabidopsis. Our results, in combination with several known expressed sequences tags (ESTs), indicate that the Arabidopsis genome contains at least 12 PME-related genes. Consequently, a method of systematic gene expression analysis has been applied in order to discern the expression pattern of these 12 genes throughout the plant at the floral stage. Whereas most of these genes appeared to be more or less ubiquitously expressed throughout the plant, several genes are distinguishable by their strikingly specific expression in certain organs. The present data bring a new insight into the role of specific PME-related genes in flower and root development. 相似文献
4.
The soluble polypeptides from Cylindrotheca fusiformis were labelled with [35S]O42− and resolved by two-dimensional gel electrophoresis. More than 600 polypeptides were detected upon a 26-day exposure to X-ray film. Analysis of the labelling pattern during the cell cycle show that labelling of at least 208 polypeptides changes; the majority, however, remain unchanged. Most of the changes occur in the beginning of the cell cycle and typically involve increases; those occurring in the second half of the cycle typically involve decreases. Light or its absence affects apparent protein turnover and the labelling rates of several polypeptides. Polypeptide labelling during the cell cycle was used as a reference to analyse the effect of silicate deprivation on diatom metabolism. In the absence of silicate, protein turnover increases: however, the addition of silicate counteracts but does not fully reverse this change. Silicate starvation affects the program of synthesis for several polypeptides, but in general the program of polypeptide labelling continues up to the S phase of the cell cycle. Addition of silicate to silicate-starved cells causes the appearance of four hitherto undetected polypeptides. 相似文献
5.
硝态氮是作物吸收无机氮素的主要形态,硝酸盐转运蛋白2(nitrate transporter 2,NRT2)作为高亲和性的转运蛋白,以硝酸盐作为特异性底物,在可利用的硝酸盐受限时,高亲和性转运系统被激活,在硝酸盐吸收、转运过程中发挥着重要作用。大多数NRT2不能单独转运硝酸盐,需在硝酸盐同化相关蛋白2(nitrate assimilation related protein 2,NAR2)的协助下才能完成硝酸盐的吸收或转运。作物氮利用效率受环境条件影响,品种间存在差异,因此培育高氮素利用效率品种有重大意义。高粱(Sorghum bicolor)具有耐贫瘠特性,对土壤中的氮素吸收和利用效率较高。本研究结合高粱基因组数据库对NRT2/3基因家族成员基因结构、染色体定位、理化性质、二级结构与跨膜结构域、信号肽与亚细胞定位、启动子区顺式作用元件、系统进化、单核苷酸多态性(single nucleotide polymorphism,SNP)的识别与注释及选择压力进行了全面分析。通过生物信息学分析,筛选出5个NRT2s(命名为SbNRT2-1a、2-1b、SbNRT2-2–4)基因和2个NAR2s(SbNRT3-1–2)基因,较谷子略少。分布在3条染色体上,分为4个亚家族,同一亚族中基因结构高度相似;高粱NRT2/3亲水性平均值均为正值,表明均为疏水性蛋白;α-螺旋和无规则卷曲占二级结构总量的比例大于70%;亚细胞定位均在质膜上,其中NRT2s蛋白不含信号肽,NRT3s蛋白含信号肽;进一步对其跨膜结构域进行分析,发现NRT2s家族成员跨膜结构域个数均大于10个,而NRT3s家族成员跨膜结构域个数为2个;高粱与玉米(Zea mays)NRT2/3s的共线性较好;蛋白结构域显示存在MFS_1和NAR2蛋白结构域,可执行高亲和力硝酸盐转运;系统进化树分析可知,高粱与玉米和谷子的NRT2/3基因亲缘关系更近;基因启动子顺式作用元件分析发现,SbNRT2/3基因的启动子区均具有数个植物激素和逆境应答元件,可以响应高粱生长和环境变化;基因表达热图显示低氮条件下在根诱导表达的是SbNRT2-1a、SbNRT2-1b和SbNRT3-1,推测可在高粱根部表达并调控对硝酸盐的吸收或转运过程。在SbNRT2-4和SbNRT2-1a等发现多个非同义SNP变异;选择压力分析表明,高粱NRT2/3基因家族在进化过程中受纯化选择作用。SbNRT2/3基因表达及蚜虫侵染影响与基因在不同组织中的表达分析结果一致,SbNRT2-1b和SbNRT3-1在感染蚜虫品系5-27sug根部表达显著,高粱蚜虫侵染叶片显著降低了SbNRT2-3、SbNRT2-4和SbNRT3-2的表达水平。本研究初步对高粱全基因组NRT2/3基因家族进行鉴定、表达与DNA变异分析,为高粱氮高效研究提供了基础。 相似文献
6.
磷转运蛋白1(phosphate transporter protein 1, PHT1)家族在植物对磷的吸收及再利用过程中发挥重要作用。该研究对菠萝PHT1基因(AcoPHT1)进行全基因组鉴定,并对基因结构、编码蛋白保守功能域和基因表达进行了分析。结果表明:(1)共鉴定到9个AcoPHT1基因,位于基因组7个连锁群上,所有基因均含有1~3个内含子,内含子相位类型多样。(2)除AcoPHT1.8外,AcoPHT1蛋白均为碱性蛋白,所有蛋白属于亲水性蛋白,且含有10~13个跨膜功能域,均具有保守的PHT1蛋白标签序列GGDYPLSATIxSE,主要定位于叶绿体和细胞质中。(3)AcoPHT1蛋白聚类在单子叶植物组和单双子叶植物混合组中,相对于拟南芥,水稻PHT1与菠萝PHT1相似度更高。(4)AcoPHT1基因启动子区含有P1BS、W-box等与磷吸收和响应胁迫有关的多个顺式作用元件。(5)靶基因预测分析显示,基因AcoPHT1.2、AcoPHT1.8和AcoPHT1.9受多个miRNA调控。(6)AcoPHT1基因表达存在组织特异性和功能冗余性,不同PHT1基因可能在菠萝不同组织或发育阶段发挥作用。该研究结果为菠萝PHT1家族基因的功能鉴定和育种应用奠定理论基础。 相似文献
7.
OsPT6:1, a phosphate transporter encoding gene from the leaf samples of Oryza sativa, was identified through PCR with specifically designed primers. The phylogenetic analysis and the conserved amino acid residue
site detection suggested OsPT6:1 a possible high-affinity phosphate transporter encoding gene. In situ hybridization and RT-PCR demonstrated the expression of OsPT6:1 in both roots and leaves. The peak expression signal was observed in mesophyll cells under low phosphorus (P) induction.
A homologous recombination study indicated that OsPT6:1 can enhance the Pi uptake efficiency of Pichia pastoris. At the meantime, the introduction of OsPT6:1 was able to complement the Pi uptake function of yeast cells with high-affinity phosphate transporters deficient. Those results
substantiated our contention that OsPT6:1 encoded a high-affinity phosphate transporter of Oryza sativa. 相似文献
8.
高亲和性K+转运蛋白(high-affinity K+ transporter,HAK)是植物中最重要的K+转运蛋白家族之一,在植物K+吸收和转运过程中发挥重要功能。为探究甜菜BvHAK基因家族成员生物学功能及基因表达模式,采用生物信息学手段,预测了蛋白质的理化性质、基因结构、染色体定位、系统进化、保守基序、三维结构、互作网络、启动子顺式作用元件,并通过qRT-PCR分析了盐胁迫下BvHAKs基因在甜菜不同组织中的表达水平。共鉴定出10个甜菜BvHAK基因家族成员,含有8-10个外显子、7-9个内含子;平均氨基酸个数为778.30,平均分子量为88.31 kDa,等电点为5.38-9.41,跨膜区为11-14个。BvHAK4、-5、-7和-13定位在质膜,而其余定位在液泡膜。系统进化分析发现,高等植物HAK可分为5个簇,分别为Ⅰ、Ⅱ、Ⅲ、Ⅳ和Ⅴ簇,其中Ⅱ簇成员可进一步分为Ⅱa、Ⅱb和Ⅱc等3个亚簇;BvHAK家族成员则分布在前4簇,分别含有1、6、1和2个成员。甜菜BvHAK基因家族主要含有胁迫响应元件、激素响应元件和生长发育响应元件。进一步对BvHAK基因在盐处理下甜菜不同组织中的表达模式分析发现,50和100 mmol/L NaCl不同程度地诱导甜菜地上部和根部BvHAK基因家族成员的表达;高盐(150 mmol/L)则下调了其在地上部的表达水平。这些结果表明,BvHAK基因家族在响应盐胁迫过程中起重要作用。 相似文献
9.
苯丙氨酸解氢酶(phenylalanine ammonia-lyase,PAL)基因家族参与苯丙烷类代谢过程,通过调控植物抗病次生物质的合成在植物抗逆反应中发挥重要作用。为明确谷子PAL基因家族在逆境胁迫下的表达规律,该研究利用生物信息学方法对谷子PAL基因家族进行鉴定和表达分析。结果表明:谷子具有11个PAL基因,在进化树中可分为3个亚家族,SiPAL7独自进化为一支。通过构建蛋白结构域发现PAL基因家族成员均含有保守的PAL结构域。启动子分析显示,PAL基因含有应答激素、逆境胁迫等多种因子的顺式作用元件,说明PAL基因广泛参与不同生物学调控过程。RT-qPCR结果显示,谷子PAL基因家族多为诱导型表达,不同光照条件下PAL基因表达量变化明显,不同基因具有不同响应模式,说明谷子PAL基因家族在参与光调节反应中发挥重要作用。谷子PAL基因高度保守,广泛响应不同非生物胁迫,具有表达特异性。该研究结果为揭示PAL基因家族在调节谷子抗性及胁迫应答过程中的作用提供了参考。 相似文献
10.
TIFY基因家族在茶树激素信号转导及其逆境胁迫具有十分重要的作用。该文利用生物信息学方法对茶树基因组中的TIFY家族进行了鉴定,分析其理化性质、系统进化、基因结构、染色体定位、启动子区顺式作用元件、组织表达模式,并通过RT-qPCR对部分TIFY家族成员进行了非生物胁迫。结果表明:(1)茶树TIFY基因家族成员19个(CsTIFY1~CsTIFY19),分属于4个蛋白亚家族TIFY、JAZ、ZML 和 PPD,且不均匀地分布在8 条染色体上,按照进化关系及结构特点可分为 7 组,每组内具有相似的基因结构与保守基序组成。(2)CsTIFYs基因的启动子区具有多种包含激素和非生物胁迫响应的顺式作用元件,通过实时荧光定量(RT-qPCR)分析其家族成员对在茉莉酸甲酯、盐(20%氯化钠)、冷(4 ℃)以及干旱(20% PEG-6000)处理下反应强烈,部分基因在根与顶芽中有较高的表达量。由此推测,TIFY基因家族可能在茶树激素信号调控、胁迫响应、生长和发育等多方面发挥功能作用。 相似文献
11.
Komatsuzawa H Choi GH Fujiwara T Huang Y Ohta K Sugai M Suginaka H 《FEMS microbiology letters》2000,188(1):35-39
We identified a gene from Staphylococcus aureus, flp (fmtA-like protein), encoding a protein of 489 amino acid residues with a molecular mass of 56.4 kDa. The deduced amino acid sequence shows similarity to previously characterized penicillin binding proteins (PBPs) and FmtA of S. aureus (one of the factors which affect methicillin resistance). FLP protein has three motifs, which are conserved in PBPs and beta-lactamases, suggesting that it might be associated with cell wall synthesis. Recombinant FLP protein, however, lacks penicillin binding activity, and the inactivation of flp in two methicillin-resistant S. aureus strains did not cause a reduction in the methicillin resistance. 相似文献
12.
SUN基因是调控植物生长发育的关键基因。本研究鉴定了二倍体森林草莓(Fragaria vesca)的SUN基因家族,并对各成员的理化性质、基因结构、系统进化以及基因表达进行了分析。结果表明,森林草莓有31个FvSUN基因,其编码蛋白可聚类为7个组,同一组内成员具有高度相似的基因结构与编码蛋白保守域;FvSUNs蛋白的亚细胞定位主要在细胞核中。共线性分析表明森林草莓FvSUNs基因家族主要通过染色体片段复制产生,拟南芥与森林草莓存在23对直系同源基因。利用森林草莓的转录组数据,对FvSUNs基因的组织表达特征进行分析,发现主要可归为3类:各组织均表达、组织中几乎不表达、组织特异性表达,并通过实时荧光定量PCR (quantitative real-time polymerase chain reaction, qRT-PCR)进一步验证结果。此外,还对森林草莓进行不同的逆境胁迫处理,qRT-PCR分析了31个FvSUNs基因的表达情况,发现大部分基因均在不同程度上受低温、高盐或干旱胁迫的诱导表达。这些研究结果为深入揭示草莓SUN基因的生物学功能及其分子机制奠定了基础。 相似文献
13.
In this study, we performed cloning and expression analysis of six putative sucrose transporter genes, designated TcSUT1, TcSUT2, TcSUT3, TcSUT4, TcSUT5 and TcSUT6, from the cacao genotype ‘TAS-R8’. The combination of cDNA and genomic DNA sequences revealed that the cacao SUT genes contained exon numbers ranging from 1 to 14. The average molecular mass of all six deduced proteins was approximately 56 kDa (range 52 to 66 kDa). All six proteins were predicted to exhibit typical features of sucrose transporters with 12 trans-membrane spanning domains. Phylogenetic analysis revealed that TcSUT2 and TcSUT4 belonged to Group 2 SUT and Group 4 SUT, respectively, and the other TcSUT proteins were belonging to Group 1 SUT. Real-time PCR was conducted to investigate the expression pattern of each member of the SUT family in cacao. Our experiment showed that TcSUT1 was expressed dominantly in pods and that, TcSUT3 and TcSUT4 were highly expressed in both pods and in bark with phloem. Within pods, TcSUT1 and TcSUT4 were expressed more in the seed coat and seed from the pod enlargement stage to the ripening stage. TcSUT5 expression sharply increased to its highest expression level in the seed coat during the ripening stage. Expression pattern analysis indicated that TcSUT genes may be associated with photoassimilate transport into developing seeds and may, therefore, have an impact on seed production. 相似文献
14.
15.
【背景】几丁质是真菌细胞壁的重要成分,由几丁质合成酶(chitin synthase,CS)催化合成。几丁质合成酶编码基因在大型食用真菌金针菇中的数量及表达规律尚不明确。【目的】探究几丁质合成酶基因在金针菇中存在的数量及其在子实体不同发育时期的表达规律,为其在大型真菌子实体生长发育过程中的功能研究提供基础。【方法】基于已有的金针菇菌株L11基因组数据,结合NCBI其他真菌CS序列鉴定金针菇中几丁质合成酶编码基因的数量,并对其进行生物信息学分析。进一步根据金针菇F19转录组数据以及实时荧光定量PCR (RT-qPCR)技术分析金针菇CS基因家族的表达规律。【结果】在金针菇单核体菌株L11的基因组中鉴定到9个几丁质合成酶基因,系统发育分析表明它们在子实体发育过程中的表达模式可分为4类(皮尔森相关系数=0.85)。【结论】金针菇CS基因家族表达模式在金针菇不同生长发育时期均存在差异,可能参与了子实体发育不同时期和组织的形态建成。 相似文献
16.
磷脂酶D(PLD)是植物生长和胁迫反应过程中参与膜磷脂分解代谢的关键酶。PLD编码基因在高等植物中构成了一个大的基因家族,但是仍未对高粱PLD基因家族进行深入研究。本研究中,通过全基因组分析,鉴定了15个PLD基因,并分成6个亚组,初步揭示了SbPLDs的基因结构、保守结构域、染色体定位和系统发育关系等信息。基因复制的研究表明,片段复制在高粱PLD基因家族的扩展中发挥了重要作用,SbPLDs在进化过程中经历了强烈的纯化选择。此外,转录组数据表明,受启动子区上游顺式元件调控的PLD家族基因可能在高粱的生长发育中发挥重要作用。通过real-time PCR分析,显示部分SbPLDs参与非生物胁迫和激素途径的潜力。亚细胞定位表明,高粱PLD蛋白在细胞质中富集,可能具有抗逆胁迫的功能。本研究为了解高粱PLD基因的特征提供了理论参考,为研究高粱PLD基因家族的进化提供了新的视角,也为阐明高粱PLD基因家族的功能提供了基础信息。 相似文献
17.
Peelman F Labeur C Vanloo B Roosbeek S Devaud C Duverger N Denèfle P Rosier M Vandekerckhove J Rosseneu M 《Journal of molecular biology》2003,325(2):259-274
An alignment of the mammalian ABCA transporters enabled the identification of sequence segments, specific to the ABCA subfamily, which were used as queries to search for eukaryotic and prokaryotic homologues. Thirty-seven eukaryotic half and full-length transporters were found, and a close relationship with prokaryotic subfamily 7 transporters was detected. Each half of the ABCA full-transporters is predicted to comprise a membrane-spanning domain (MSD) composed of six helices and a large extracellular loop, followed by a nucleotide-binding domain (NBD) and a conserved cytoplasmic 80-residue sequence, which might have a regulatory function. The topology predicted for the ABCA transporters was compared to the crystal structures of the MsbA and BtuCD bacterial transporters. The alignment of the MSD and NBD domains provided an estimate of the degree of residue conservation in the cytoplasmic, extracellular and transmembrane domains of the ABCA transporter subfamily. The phylogenic tree of eukaryotic ABCA transporters based upon the NBD sequences, consists of three major clades, corresponding to the half-transporter single NBDs and to the full-transporter NBDls and NBD2s. A phylogenic tree of prokaryotic transporters and the eukaryotic ABCA transporters confirmed the evolutionary relationship between prokaryotic subfamily 7 transporters and eukaryotic ABCA half and full-transporters. 相似文献
18.
Na+/H+逆向转运蛋白(Na+/H+antiporter,NHX)基因家族在植物响应盐胁迫中发挥重要作用。本研究鉴定了大白菜NHX基因家族成员,并分析了大白菜NHX基因(Brassica rapa ssp.Pekinensis NHX,BrNHXs)响应高温、低温、干旱和盐胁迫等非生物逆境的表达模式。结果表明,在大白菜中共鉴定到9个NHX基因家族成员,分布在大白菜的6条染色体上,其氨基酸数目在513–1154 aa之间,相对分子量集中在56804.22–127856.66 kDa,等电点位于5.35–7.68之间。该基因家族成员主要存在于液泡中,基因结构完整,外显子的数目介于11–22之间。大白菜NHX基因家族编码的蛋白质二级结构都具有α-螺旋、β-转角和不规则卷曲结构,其中α-螺旋发生频率较高。实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)分析显示,该基因家族成员在高温、低温、干旱和盐胁迫下均有不同程度地响应,且在不同时间表达差异显著。以BrNHX02和BrNHX09对这4种胁迫的响应最为显著,表达量在处理72 h时均显著上调,可作为候选基因进一步验证其功能。 相似文献
19.
WRKY是植物特有的一个转录因子超家族,参与植物生长发育、物质代谢以及生物和非生物胁迫响应等多种生物学过程的调控。尽管WRKY转录因子基因已在高等植物广泛表征,然而有关真核藻类WRKY却知之甚少。采用多序列比对、系统进化和保守域分析等技术对30种真核藻类WRKY进行全基因组鉴定,共获得24个WRKY成员,均来自绿藻门(Chlorophyta)藻类。红藻门(Rhodophyta)、灰藻门(Glaucophyta)和硅藻门(Bacillariophyta)等藻类未检测到WRKY。24个WRKY成员均具有保守结构域七肽序列WRKYGQ(E/A/H/N)K和锌指基序C-X4-5-C-X22-23-H-X-H,分别归类于Ⅰ、Ⅱa、Ⅱb和R组。高含虾青素的雨生红球藻(Haematococcus pluvialis)含有2个Ⅰ类WRKY成员(HaeWRKY-1和HaeWRKY-2)。进一步克隆HaeWRKY-1和HaeWRKY-2基因编码序列,构建原核表达载体在大肠杆菌BL21(DE3)中诱导表达,并通过Ni-NTA亲和层析获得纯化的HaeWRKY融合蛋白。雨生红球藻在正常培养条件下,HaeWRKY-1表达量显著高于HaeWRKY-2。高光逆境胁迫显著上调HaeWRKY-1表达和下调HaeWRKY-2表达。HaeWRKY基因启动子含有多个光、乙烯、脱落酸(abscisic acid, ABA)以及逆境响应顺式元件。特别在HaeWRKY-2启动子区未检测到W-box顺式元件,但HaeWRKY-1和控制虾青素生物合成关键酶基因HaeBKT、HaePSY基因启动子含有W-box元件。基于本研究及前人的发现,我们推测高光胁迫下HaeWRKY-2低表达可能导致HaeWRKY-1的上调表达,HaeWRKY-1进而上调虾青素合成关键基因(HaeBKT、HaePSY等)表达,促进虾青素合成积累。这为深入解析雨生红球藻高光胁迫响应及虾青素合成的调控机制提供了新思路。 相似文献
20.
Dicer proteins belong to the RNase III family of proteins, which are key components in small RNA biogenesis. In Solanum lycopersicum, seven Dicer-like (DCL) genes have been identified and have been named SlDCL. In this study, we cloned the full-length sequence of the SlDCL genes including untranslated regions using RNA ligase-mediated rapid amplification of cDNA ends. Our analysis indicates that 7 SlDCLs were located on 5 tomato chromosomes (6, 7, 8, 10, and 11). The gene structure of the SlDCLs covered long genomic regions and contained more than 20 exons. Phylogenetic analysis divided the seven SlDCL members into four subgroups. In general, all seven SlDCLs were expressed in all organs but more in flowers and fruits than in the other parts. Moreover, the expressions of some genes changed slightly after treatment with ethylene or 1-methylcyclopropene suggesting their likely roles in plant responses to ethylene. Our findings provide essential information on SlDCL genes in tomato and will aid in the functional classification of DCL families in plants. 相似文献