Genomic Analysis of MicroRNA Promoters and Their Cis-Acting Elements in Soybean

MicroRNAs （miRNAs） are derived from distinct loci in the genome and play crucial roles in RNA-mediated gene silencing mechanisms that regulate cellular processes during development and stress responses of plants. The miRNAs are approximately 21 nucleotides long and code for the complementary strand to a larger genic mRNA. They are often found within the complementary primary transcript （pri-miRNAs）. In the past few years, a growing number of soybean miRNAs have been discovered, however, little is known about the transcriptional regulation of these miRNAs. In this study, promoters and cis-acting elements of soybean miRNAs were analyzed using the genomic data for the first time. A total of 82 miRNAs were located among 122 loci in genome, some were present as double or multiple copies. Five clusters that included ten miRNAs were found in genome, and only one cluster share the same promoter. A total of 191 promoters from 122 loci of the soybean miRNA sequences were found and further analyzed. The results indicated that the conserved soybean miRNA genes had a greater proportion of promoters than that of non-conserved ones, and the distribution of the transcript start sites （TSSs） and TATA-boxes found had different motif styles between conserved and non-conserved miRNA genes. Furthermore, the cis-acting elements 5＇ of the TSSs were analyzed to obtain potential function and spatiotemporal expression pattern of miRNAs. The data obtained here may lead to the identification of specific sequences upstream of pre-miRNAs and the functional annotation of miRNAs in soybean.

猪嵌合RNA BCL2L2-PABPN1剪接调控、表达与亚细胞定位分析

相邻基因间顺式剪接(cis-splicing of adjacent gene,cis-SAGe)产生的嵌合RNA在细胞生长、疾病发生等生理活动中发挥重要作用,为探究cis-SAGe剪接调控机制,研究在前期克隆BCL2L2-PABPN1(BP)基础上,利用minigene、定点突变、瞬时转染、半定量RT-PCR以及生物信息学分析等方法分析BP转录后剪接的调控元件,发现thyroid hormone receptor exon 9、gh-1 intron 3、srp40-exonic splicing enhancer、β-tropomyosin exon 6b等剪接增强子元件在嵌合子形成中发挥重要作用。同时比较分析BP与两个亲本基因组织表达以及亚细胞定位情况。研究结果将为进一步揭示BP转录后调控机制及功能提供基础。

Isolation of cDNAs Enconding Two Distinct Transcription Factors Binding to DRE cis-acting Element Involved in Cold-and drought-induced Expression of Arabidopsis rd29A Gene

Previous study has defined DRE (dehydration responsive element) cis acting element and its important role in expressions of Arabidopsis rd29A gene under cold, dehydration and high salt stresses. In order to clarify the expression mechanism of rd29A gene, we isolated two cDNA clones that encoded DRE binding proteins ( DREB1 and DREB2 ) from cold and drought treated Arabidopsis plants, using DRE cis acting element in the promoter region of rd29A gene and yeast One Hybrid screening method. Experiments showed both DREB1 and DREB2 specifically interacted with DRE cis element. Homologous analysis showed no significant similarity between DREB1 and DREB2 in whole deduced amino acid sequences. However, both DREB1 and DREB2 proteins contained a conserved DNA binding domain (AP2/EREBP domain). Structural analysis of proteins also showed they had a nuclear localization signal (NLS) in their N terminal region and an acidic activation region in their C terminal region. AP2/EREBP domain is composed of 58 amino acids, which presents in a large family of plant genes encoding DNA binding proteins. We analyzed many plant transcription factors containing conserved AP2/EREBP domains. The 14th valine (V) and 19th glutamate (E) in the amino acid sequence of AP2/EREBP domains might be the consensus recognizing and binding to DRE cis element. Northern analysis indicated that DREB1 gene was induced by low temperature, whereas DREB2 gene was induced by dehydration and high salt stresses. Present studies suggest that the expression of rd29A gene under low temperature, dehydration and high salt stresses is regulated by DREB1 and DREB2 transcriptional factors in two separate signal transduction pathways, respectively.

Isolation and analysis of a TIR-specific promoter from poplar

A 5＇ flanking region of the well-conserved Toll/interleukin-1 receptor domain （TIR）-encoding sequence was isolated from the genomic DNA ofMelampsora magnusiana Wagner resistant clones of hybrid triploid poplars [（Populus tomentosa × P. bolleana） × P. tomentosa]. Sequencing results and alignment analysis show that the obtained TIR-specific promoter （named as PtTIRp01） was 1,732 bp in length; moreover 3＇ region of the PtTIRp01 contains a responds to the 5＇ composition of TIR-NBS type gene PtDRG02, of 747 bp long 5＇ region of TIR-NBS type gene PtDRG02 and its 398 bp complete TIR-encoding sequence, which significantly corindicating that the obtained TIR-specific promoter region consists upstream region of promoter （985 bp）. It was found that the 5＇ region of TIR-NBS type gene PtDRG02 was characterized in the downstream region of the transcriptional start, named as 5＇-untranslated region （5＇ UTR）, consisting of one 93 bp 5＇-untranslation exon, one 213 bp intron and one 441 bp TIR-encoding open reading frame （ORF）. In addition, several putative cis-acting motifs were present in the obtained TIR-specific promoter of PtDRG02, including one TATA box, one GC-rich, one AT-rich, one P-box, one 3-AF1 binding site, two CAAT boxes, two GT-1 motifs, three typical W-boxes, four I-boxes, and one multi-cis-acting fragment （MCF）. The latter contains five types of regulatory elements （E4, G-box, ABRE motif, box 1 and HVA 1 s）, most of which were homologous to the cis-acting regulatory elements involved in the activation of defense genes in plants. Thus, it can be suggested that TIR-specific promoter might be a pathogen-inducible promoter and be necessary for the inducible expression of defense-related genes. Key words Toll/interleukin- 1 receptor domain, promoter, Cis-acting element, poplar

Cis-acting regulatory elements： from random screening to quantitative design

The cis-acting regulatory elements, e.g., promoters and ribosome binding sites （RBSs） with various desired properties, are building blocks widely used in synthetic biology for fine tuning gene expression. In the last decade, acquisition of a controllable regulatory element from a random library has been established and applied to control the protein expression and metabolic flux in different chassis cells. However, more rational strategies are still urgently needed to improve the efficiency and reduce the laborious screening and multifaceted characterizations. Building precise computational models that can predict the activity of regulatory elements and quantitatively design elements with desired strength have been demonstrated tremendous potentiality. Here, recent progress on construction of cis- acting regulatory element library and the quantitative predicting models for design of such elements are reviewed and discussed in detail.

GCC box in Arabidopsis PDF1.2 promoter is an essential and sufficient cis-acting element in response to MeJA treatment

The expression of Arabidopsis PDF1.2 gene is regulated by jasmonic acid (JA) and ethylene (ET). It also has been well documented that GCC box is an element re- sponsive to ET,however, the responsive mechanism of JA in such plant defense gene expression is unclear. In this paper, the authors define the essential cis-acting element in PDF1.2 promoter responsive to methyl jasmonate (MeJA) through fragment deletions and site-directed mutageneses combining Agrobacterium-mediated transient reporter gene expression in tobacco leaves. Firstly, the MeJA inducible expression of PDF1.2 was confirmed by using the upstream ?1.86 kb fragment of PDF1.2 gene. Secondly, the upstream –300— ?243 bp fragment of the promoter was evidenced to respond to MeJA. To further characterize this promoter region, three point mutations were introduced into the –300—?243 bp fragment of the promoter. This result showed that the muta- tion of GCC box abolished MeJA induction, whereas the mutations of the G box-like and the imperfect palindrome sequence did not significantly decrease MeJA inducible effect, indicating that GCC box in PDF1.2 is essential for MeJA induction. The sufficient responsiveness to MeJA of this GCC box was further investigated by 4×GCC fused up- stream to the CaMV 35S minimal promoter. This result sug- gested that the fused promoter was able to activate reporter gene expression in response to MeJA. Thus these results in- dicate that the GCC box in PDF1.2 is an essential and suffi- cient element to confer MeJA induction.

MicroRNA Primary Transcripts and Promoter Elements Analysis in Soybean (Glycine max L. Merril.)

The importance of microRNA （miRNA） at the post-transcriptional regulation level has recently been recognized in both animals and plants. In recent years, many studies focused on miRNA target identification and functional analysis. However, little is known about the transcription and regulation of miRNAs themselves. In this study, the transcription start sites （TSSs） for 11 miRNA primary transcripts of soybean from 11 miRNA loci （of 50 loci tested） were cloned by a 5＂ rapid amplification of cDNA ends （5＂ RACE） procedure using total RNA from 30-d-old seedlings. The features consistent with a RNA polymerase II mechanism of transcription were found among these miRNA loci. A position weight matrix algorithm was used to identify conserved motifs in miRNA core promoter regions. A canonical TATA box motif was identified upstream of the major start site at 8 （76%） of the mapped miRNA loci. Several cis-acting elements were predicted in the 2 kb 5＂ to the TSSs. Potential spatial and temporal expression patterns of the miRNAs were found. The target genes for these miRNAs were also predicted and further elucidated for the potential function of the miRNAs. This research provides a molecular basis to explore regulatory mechanisms of miRNA expression, and a way to understand miRNA-mediated regulatory pathways and networks in soybean.

中华猕猴桃全基因组MADS-box基因家族鉴定及表达分析

【目的】鉴定并分析中华猕猴桃MADS-box基因家族成员,探明MADS-box基因家族成员在果实生长发育及芽休眠中的表达模式。【方法】基于中华猕猴桃红阳V3基因组数据库,利用生物信息学方法对中华猕猴桃MADS-box全基因组进行鉴定,分析其家族成员的理化性质、系统进化树、保守基序、顺式作用元件,并对其在果实生长发育及不同组织中的表达模式进行分析。【结果】在中华猕猴桃红阳基因组中鉴定到了68个AcMADS-box基因,共包含11个亚家族,不均匀地分布于22条染色体上,成员间共存在32对共线性基因对;在基因家族上游启动子区域发现与光响应、激素响应、逆境胁迫响应等相关的顺式元件;17个AcMADS-box基因在组织间高表达且有表达特异性,推测是参与调控中华猕猴桃生长发育的关键基因。【结论】初步鉴定并提供了AcMADS-box家族成员信息,16个AcMADS-box家族成员在根、枝、茎、叶、花中高表达,8个家族成员在花芽中高表达。结果为进一步研究AcMADS-box参与中华猕猴桃的生长发育调控机制提供参考。

A novel cold-inducible promoter,PThCAP from Tamarix hispida,confers cold tolerance in transgenic Arabidopsis thaliana

Our previous studies have revealed that the Th CAP gene plays a vital role in transgenic Populus(P.davidiana 9 P.bolleana) in response to cold stress.However,the regulatory mechanism of Th CAP gene expression has been unclear.In this study,the 50 flanking region of the Th CAP promoter(PTh CAP) was cloned using a genomewalking method.By analyzing cis-acting regulatory elements of PTh CAP,a DRE motif and MYC and MYB elements were found to be located in the promoter.To identify the regulatory elements that control the expression of the Th CAP gene promoter,a series of deletion derivatives ofPTh CAP,P1–P5,from the translation start code(-1538,-1190,-900,-718 and-375 bp),were fused to the GUS reporter gene,and then each deletion was stably introduced into Arabidopsis thaliana plants.Deletion analysis of the promoter suggested that only the P2 fragment had strong GUS expression in leaves and roots of A.thaliana exposed to low temperature stress.These results suggest that this290-bp region(-1190 to-900 bp),as an important part in PTh CAP,was associated with cold tolerance of A.thaliana.Our results provide evidence for the regulatory mechanism of Th CAP gene involved in the response to cold stress,and that the gene is promising candidate gene for genetic improvement of crops.

番茄SlLCY-B2及其启动子的分子特征和sgRNA分析

【目的】番茄红素合成后会在β-番茄红素环化酶SlLCY-B(lycopene-β-cyclase)催化下进入代谢途径,抑制SlLCY-B2的表达可增加番茄红素的积累。对番茄红素代谢调控基因SlLCY-B1的分子特征及sgRNA进行系统分析,为利用CRISPR/Cas9技术对SlLCY-B2及其上游启动子序列进行定点突变或片段删除、调控SlLCY-B2的表达以减少代谢的途径,提高番茄果实的番茄红素含量。【方法】利用生物在线工具,对SlLCY-B2多肽特性及保守结构域、基因组结构、数字表达谱、启动子顺式作用元件进行系统分析,根据CRISPR-Cas9靶点设计原则,设计筛选出适宜的sgRNA。【结果】SlLCY-B2位于番茄6号染色体,编码498个氨基酸,gDNA不含内含子,呈果实特异性表达。SlLCY-B2分布145条sgRNA,其中21条有高特异性。SlLCY-B2上游-1500 bp启动子序列分布15个顺式作用元件和112条sgRNA。【结论】对SlLCY-B2和其启动子区进行基因编辑,可调控SlLCY-B2的表达,将提高番茄红素含量。

牡丹花青素苷合成关键基因PsDFR启动子活性分析

[目的]分析牡丹花青素苷合成关键基因PsDFR启动子的顺式作用元件及活性,为进一步研究PsDFR启动子的功能及其在牡丹花色形成中的调控机制奠定基础。[方法]以‘黑花魁’牡丹花瓣中提取的基因组DNA为模板,通过染色体步移法克隆PsDFR的启动子序列。利用生物信息在线软件预测分析启动子序列的顺式作用元件。构建5个不同长度缺失启动子与GUS基因融合的表达载体,并瞬时转化烟草叶片,通过GUS组织化学染色和酶活性检测分析缺失启动子的活性,及其对脱落酸(ABA)、茉莉酸甲酯(MeJA)、光照等不同胁迫处理的响应。[结果]克隆得到PsDFR上游1687 bp的启动子序列。生物信息学分析发现PsDFR启动子含有多个光信号、激素响应、逆境响应及组织特异表达等顺式作用元件,这暗示PsDFR的表达可能受光信号、激素和胁迫等多种信号的共同调节。GUS组织化学染色和酶活性检测表明,随启动子片段的缩短,启动子活性逐渐下降,-1623至-916区段对于启动子活性具有重要作用。MeJA和黑暗处理对各启动子片段活性均有显著抑制作用,恢复光照可促使启动子活性明显回升,而参与ABA响应的核心调控区域位于-443至-76 bp。[结论]PsDFR启动子含有多个光信号、激素响应、逆境响应及组织特异表达等顺式作用元件,其活性受光的正调控以及MeJA的负调控;-1623至-916 bp间的区域对于启动子活性具有重要作用,-443至-76 bp是响应ABA处理的核心区域。该研究为进一步揭示PsDFR应答环境信号参与牡丹花色形成的分子调控机制提供参考。

桑树NAC基因家族鉴定与生根粉处理下表达模式分析

【目的】探究桑树NAC基因家族的生物信息学特征,并研究1000 mg·L^(-1)生根粉(ABT)处理对该基因产生的影响,为深入研究桑树NAC转录因子的功能提供依据。【方法】利用生物信息学方法对川桑NAC基因家族进行基因鉴定,并分析其理化性质、保守基序、顺式作用元件、系统进化树、蛋白三级结构及互作关系,并选用‘果桑大十’作为试验材料,经ABT处理后,分析其4个时期(10、20、30、40 d)NAC基因家族的表达模式。【结果】桑树NAC基因家族共有92个成员,分为22个亚家族,亚家族成员广泛分布于细胞核,且具有相似的保守结构。顺式作用元件分析表明,桑树NAC基因家族对激素具有较强的响应能力。蛋白三级结构显示同一亚家族的蛋白空间结构相似,且功能相同,这可能与其进化同源性有关。ABT处理后,NAC基因家族的表达水平整体呈上升趋势,而MnNAC91、MnNAC67、MnNAC28在对照组(CK)中呈现较高的表达水平。【结论】NAC基因家族功能相对稳定,在整个进化过程中无明显变化;ABT可促进NAC基因家族表达,尤其是MnNAC75、MnNAC104、MnNAC7、MnNAC30、MnNAC101、MnNAC83、MnNAC102具有较高的同源性,并与拟南芥的结构功能相似,推测对抗干旱胁迫有一定作用。

Functional analysis of cis-acting sequences regulating root-specific expression in transgenic tobacco

Two different length fragments, RSF1 and RSF2 which contained the cis-acting sequences of root-specific gene TobRB7, were isolated from tobacco genome. The abilities of these fragments to direct root-specific expression were studied by fusing them to the β-glucuronidase (GUS) report gene with different directions. After the recombined vectors were transformed into tobacco, the expression pattern was performed by histochemical staining and the quantitative analysis of GUS activity. The data suggested that the cis-acting element of TobRB7 gene direct GUS expression not only as root-specific but also as bidirectional. In our studies, the short fragment, RSF2, performed stronger activity than RSF1 with any direction. The stronger activity of GUS expression was determined by reverse inserting of RSF1 or RSF2 than positive inserting.

Developmental stage-specific factors in the mouse haematopoietic tissues binding to the 5'-flanking as-acting elements of humanε-globin gene

The human ε-globin gene is expressed in a tissue-specific and developmental stage-specific manner. During the earliest stage of gestation, this gene is expressed in the yolksac, but is silenced completely at the 6th-8th weeks of gestation. Recently, several studieson the transgenic mice have shown that the 5′-flanking DNA sequences of human

Isolation and characterization of a novel cis-acting sequences regulating root-specific gene from Daucus carota L.

Aquaporins are ubiquitous channel proteins that facilitate the transport of water across cell membranes. Most of aquaporins are represent in more than one tissues, but some of them performed highest in roots. They are be- lived to participate in water transport. DcRB7, a member of the aquaporin family, was isolated from somatic embryos of carrot and identified as a homologous gene of TobRB7. Further studies showed that the expression of DcRB7 was particular in carrot root. To investigate the transcription regulation of DcRB7, a 650-bp upstream sequence of DcRB7 was isolated by inverse PCR, and was fused to the β-glucuronidase (GUS) report gene. After the recombined vectors were transformed into tobacco, the expression pat- tern was performed by histochemical staining and the quan- titative analysis of GUS activity. The results indicated that the cis-acting element of DcRB7 gene directs GUS expression not only as root-specific but also as drought inducible.

延胡索去甲乌药碱合成酶基因CyNCS1克隆及其表达模式分析

【目的】克隆延胡索去甲乌药碱合成酶(norcoclaurine synthase, NCS)基因,分析其在延胡索异喹啉类生物碱合成中的关键作用。【方法】基于转录组数据库,利用逆转录PCR克隆延胡索CyNCS1基因及其启动子区域序列,采用生物信息学方法分析其编码蛋白的理化性质、结构特征及其启动子区域顺式作用元件,同时进行多序列比对和系统进化树分析,确定其进化关系;利用实时荧光定量PCR对其根、茎、叶及发育早、中、晚期块茎的组织表达模式进行分析;对茉莉酸甲酯(MeJA)、脱落酸(ABA)处理0,4,8,12 h的叶片进行表达谱分析,以体积分数75%酒精溶剂处理为对照(CK),确定该基因的表达特征。【结果】CyNCS1基因开放读码框长873 bp,编码291个氨基酸,编码蛋白分子量为33.09 ku,等电点为6.90,具有去甲乌药碱合成酶保守的催化结构域(GDGGVGTV/IL、YKEKF和MIEGGHLDMG),且不含信号肽,预测定位于细胞核中。多序列比对结果显示,CyNCS1与石生黄堇、罂粟的NCS蛋白同源性较高,分别为83.6%和82.1%;系统进化树结果显示,CyNCS1与石生黄堇CsNCS蛋白聚为一支,说明二者亲缘关系较近。该基因启动子区长2 238 bp,包含多种植物激素及低温、热胁迫等环境因子的顺式作用元件。实时荧光定量PCR结果显示,随着发育推进块茎中CyNCS1的表达量显著增高,且受到MeJA和ABA的诱导,MeJA处理8 h时表达量为0 h的3.10倍,ABA处理8 h为0 h的3.16倍;而对照CyNCS1表达量随时间推移无明显变化。【结论】克隆并鉴定了延胡索NCS酶基因CyNCS1,明确了其在发育进程中的表达模式,并确定该基因表达受MeJA和ABA的诱导。

烟草BTB/POZ蛋白家族的鉴定及其在抗马铃薯Y病毒中的作用

马铃薯Y病毒(potato virus Y,PVY)侵染烟草(Nicotiana tabacum)会引起烟草脉带病,造成烟叶品质下降。broad complex,tramtrack,and bric-à-brac/pox virus and zinc finger(BTB/POZ)是动植物体内广泛存在的蛋白质家族,在植物生长和发育的各个环节都起着非常重要的作用。本研究鉴定到90个烟草BTB/POZ家族蛋白,对应71个基因。在这些蛋白质的序列中,识别到9种保守基序(motif),并根据其出现顺序将BTB/POZ家族成员划分至6个亚家族。结构域分析表明,BTB/POZ同一亚家族成员的结构域具有一致性。对BTB/POZ蛋白的三维构象进行预测发现,所有亚家族成员结构都以α螺旋为主,同一亚家族成员的三维构象相似。烟草品种K326和突变体M867(抗PVY)的BTB/POZ家族基因表达模式分析显示,叶片接种PVY后,NtBTB2M、NtBTB2K、NtBTB2L、NtBTB6E基因的表达丰度显著增加,这可能与烟草M867对PVY的抗性较强有关。顺式作用元件分析显示,NtBTB2K、NtBTB2L、NtBTB2M和NtBTB6E基因的上游2000 bp区域内含有若干逆境响应相关元件,包括水杨酸响应元件、MYB结合位点等,可能和这些基因的上调表达有关。本研究为BTB/POZ蛋白的功能研究提供了理论依据,为烟草抗病育种提供了一定参考。

拟南芥RBCS-1A基因受光调节表达模式及其启动子遗传转化应用评价

RBCS编码光合碳同化关键酶核酮糖1,5-二磷酸羧化酶/加氧酶的小亚基,是控制植物光合作用的重要基因之一。本研究利用实时荧光定量PCR技术分析了拟南芥RBCS-1A受光调节的表达模式,结果表明,AtRBCS-1A表达受到光的诱导,同时具有组织表达特异性;运用生物信息学手段分析发现,该基因启动子序列中存在多个参与光应答的顺式作用元件;采用PCR技术从拟南芥基因组中分离到长度为1691bp的AtRBCS-1A启动子片段,将该片段与GUS报告基因融合构建植物表达载体并转化野生型拟南芥,对获得的转基因植株进行GUS染色,结果显示,AtRBCS-1A启动子是光诱导型和组织特异型启动子。以上结果初步证明,AtRBCS-1A启动子应用于植物遗传转化切实可行,具有重要应用价值。

Trametessp.AH28-2漆酶A的诱导合成及其基因5′-端调控区的克隆与分析

Trametessp.AH28_2漆酶同工酶的合成需要铜离子的存在,较高浓度的Cu2+有利于漆酶合成。在以葡萄糖为碳源补加0·5mmol/LCu2+的培养基中生长时,发酵液漆酶活性为44·3u/L,同时补加4·0mmol/L邻甲苯胺时,漆酶酶活提高到71·0u/L;而在补加Cu2+和邻甲苯胺的纤维二糖培养基中,酶活上升至2584u/L,为葡萄糖培养基的36·4倍。邻甲苯胺和铜离子诱导产生的漆酶同工酶组分,均为漆酶A(LacA)。竞争性RT_PCR分析表明,漆酶A基因(lacA)转录本的累积伴随有发酵液漆酶活性的增加,邻甲苯胺对lacA的调控发生在转录水平。lacA结构基因长2110bp,含有10个内含子;lacA的cDNA序列为1560bp,编码520aa的漆酶蛋白,其氨基酸序列与其它真菌漆酶具有较高的相似性。采用改进的反向PCR技术,扩增得到的lacA5′_端调控区长1881bp,分析表明,该区域上分布有1个TATA框、7个CAAT框和多个潜在的顺式作用元件序列位点,包括5个MRE元件、9个CreA结合位点、4个XRE元件、2个STRE元件和7个氮因子调控位点等。这些序列位点的存在部分地对应了菌株摇瓶发酵条件下lacA的表达规律。

金针菇L-赖氨酸合成通路基因鉴定及对不同光质的响应表达

为了从分子水平探究提高金针菇赖氨酸含量的环境因子(温度、光照、湿度、CO_2)及其作用机理,本研究首先在金针菇(Flammulina filiformis)单核体L11基因组中鉴定L-赖氨酸合成通路基因,并预测其启动子区顺式元件。根据预测结果,采用红、绿、蓝、红绿蓝复合4种光处理金针菇子实体,并分别用HPLC和定量PCR测定赖氨酸含量变化及其合成通路基因的响应表达。结果表明:金针菇赖氨酸由氨基己二酸(α-Aminoadipate,AAA)途径合成,通路中8步催化酶的编码基因及其序列在L11中全部得到鉴定。关键限速酶基因HCS、HAC1和SHD基因的启动子区存在G-box等光照响应元件。定量PCR表明,总体上看赖氨酸合成通路前7步的酶编码基因对4种光质均有响应表达。进一步比较本试验中4种不同光质的处理效果,发现蓝光照射后的金针菇子实体中,共有7个赖氨酸合成通路基因(HCS、HAC1、HAH、HDH、AAT、AAR和SR)表达量上调,同时赖氨酸含量最高。推测蓝光可能通过激活赖氨酸合成通路基因启动子区的G-box等顺式元件,诱导通路基因上调表达,进而提高了金针菇子实体中赖氨酸含量。