Roles of NAC transcription factors in cotton

Climate deterioration,water shortages,and abiotic stress are the main threats worldwide that seriously affect cotton growth,yield,and fiber quality.Therefore,research on improving cotton yield and tolerance to biotic and abiotic stresses is of great importance.The NAC proteins are crucial and plant-specific transcription factors(TFs)that are involved in cotton growth,development,and stress responses.The comprehensive utilization of cotton NAC TFs in the improvement of cotton varieties through novel biotechnological methods is feasible.Based on cotton genomic data,genome-wide identification and analyses have revealed potential functions of cotton NAC genes.Here,we comprehensively summarize the recent progress in understanding cotton NAC TFs roles in regulating responses to drought,salt,and Verticillium wilt-related stresses,as well as leaf senescence and the development of fibers,xylem,and glands.The detailed regulatory network of NAC proteins in cotton is also elucidated.Cotton NAC TFs directly bind to the promoters of genes associated with ABA biosynthesis and secondary cell-wall formation,participate in several biological processes by interacting with related proteins,and regulate the expression of downstream genes.Studies have shown that the overexpression of NAC TF genes in cotton and other model plants improve their drought or salt tolerance.This review elucidates the latest findings on the functions and regulation of cotton NAC proteins,broadens our understanding of cotton NAC TFs,and lays a fundamental foundation for further molecular breeding research in cotton.

Targeted Mutagenesis of NAC Transcription Factor Gene,OsN AC041,Leading to Salt Sensitivity in Rice

Salinity is a major abiotic stress factor that seriously affects plant growth. Many genes are involved in the response to salt stress with various metabolism pathways. A number of plant transcription factor family genes have been found to be involved in the salt stress response, and NAM, ATAF and CUC(NAC) transcription factors are thought to act as active regulators during abiotic stress, especially salt stress. In this study, we detected a rice NAC transcription factor coding gene, OsNAC041, and confirmed that it influenced the germination of seeds under salt stress and salt tolerance of plants. OsNAC041 was primarily expressed in the leaves and located in the nucleus. Furthermore, the CRISPR/Cas9 method was used to obtain a targeted osnac041 mutant, of which the plant height was higher than that of the wild-type, showing increased salt sensitivity. Moreover, RNA-seq analysis revealed a number of differentially expressed genes(DEGs) involved in several important signaling pathways in the osnac041 mutant. Subsequently, Kyoto Encyclopedia of Genes and Genomes annotation also revealed differential expression of DEGs associated with mitogen-activated protein kinase signaling, peroxisome, eukaryotictype ABC transporters, photosynthesis and plant hormones, which are involved in stress-related signaling pathways. Overall, our study suggested that OsNAC041 was involved in the salt stress response in rice. These findings not only provide empirical evidence of OsNAC041 function, but also provide new insight into its potential application in rice resistance breeding.

The NAC-like transcription factor Si NAC110 in foxtail millet(Setaria italica L.) confers tolerance to drought and high salt stress through an ABA independent signaling pathway

Foxtail millet（Setaria italica（L.）P.Beauv）is a naturally stress tolerant crop.Compared to other gramineous crops,it has relatively stronger drought and lower nutrition stress tolerance traits.To date,the scope of functional genomics research in foxtail millet（S.italic L.）has been quite limited.NAC（NAM,ATAF1/2 and CUC2）-like transcription factors are known to be involved in various biological processes,including abiotic stress responses.In our previous foxtail millet（S.italic L.）RNA seq analysis,we found that the expression of a NAC-like transcription factor,SiNAC110,could be induced by drought stress;additionally,other references have reported that SiNAC110 expression could be induced by abiotic stress.So,we here selected SiNAC110 for further characterization and functional analysis.First,the predicted SiNAC110 protein encoded indicated SiNAC110 has a conserved NAM（no apical meristem）domain between the 11–139 amino acid positions.Phylogenetic analysis then indicated that SiNAC110 belongs to subfamily III of the NAC gene family.Subcellular localization analysis revealed that the SiNAC110-GFP fusion protein was localized to the nucleus in Arabidopsis protoplasts.Gene expression profiling analysis indicated that expression of SiNAC110 was induced by dehydration,high salinity and other abiotic stresses.Gene functional analysis using SiNAC110 overexpressed Arabidopsis plants indicated that,under drought and high salt stress conditions,the seed germination rate,root length,root surface area,fresh weight,and dry weight of the SiNAC110 overexpressed lines were significantly higher than the wild type（WT）,suggesting that the SiNAC110 overexpressed lines had enhanced tolerance to drought and high salt stresses.However,overexpression of SiN AC110 did not affect the sensitivity of SiNAC110 overexpressed lines to abscisic acid（ABA）treatment.Expression analysis of genes involved in proline synthesis,Na＋/K＋transport,drought responses,and aqueous transport proteins were higher in the SiNAC110overexpressed lines than in the WT,whereas expression of ABA-dependent pathway genes did not change.These results indicated that overexpression of SiNAC110 conferred tolerance to drought and high salt stresses,likely through influencing the regulation of proline biosynthesis,ion homeostasis and osmotic balance.Therefore,SiNAC110 appears to function in the ABA-independent abiotic stress response pathway in plants.

Preliminary analysis of two NAC transcription factor expression patterns in Larix olgensis

The NAC transcription factor family is plant-specific with various biological functions.However,there are few studies on the NAC gene involving coniferous species.Bioinformatics research and expression analysis of NAC genes in Larix olgensis can be used to analyse the function of the NAC gene in the future.Screening of excellent genetic materials and molecular breeding have been utilized to cultivate high-quality,stress-resistant larches.According to the transcriptome data for L.olgensis,the genes Uni-gene81490 and Unigene70699 with complete ORFs(open reading frames)were obtained by conserved domain analy-sis and named LoNAC1 and LoNAC2,respectively.The cDNAs of LoNAC1 and LoNAC2 were 1971 bp and 1095 bp in length,encoding 656 and 364 amino acids,respectively.The molecular weights of the proteins encoded by the two genes were predicted to be 72.61 kDa and 41.13 kDa,and subcellular localization analysis indicated that the proteins were concentrated in the nucleus.The results of real-time quantitative PCR analysis showed that at different growth stages and in different tissues of L.olgensis,the relative expression levels of the two NAC genes were highest in the stem,and the expression differences were more obvious in non-lignified tissues.After drought,salt and alkali stress and hormone treatment,expression was induced to different degrees.The expression levels of LoNAC1 and LoNAC2 in semi-lignified L.olgensis were higher than in the other two periods(non-lignified and lignified),and expression levels significantly increased under drought and salt stress.Relative expression levels changed under hormone treatment.It is speculated that these two genes may not only be related to drought and salt stress and secondary growth but may also be induced by hormones such as abscisic acid.Overall,LoNAC1 and LoNAC2 are genetic materials that can be used for molecular breeding of larch.

紫花苜蓿NAC基因家族鉴定及在非生物胁迫下的表达模式分析

NAC是植物特有转录因子,在调控植物生长发育及响应非生物胁迫等方面发挥作用。为挖掘紫花苜蓿(Medicago sativa)响应干旱等非生物胁迫的NAC转录因子,本研究利用生物信息学方法在‘中苜4号’紫花苜蓿基因组中鉴定出143个NAC家族转录因子。研究表明,紫花苜蓿MsNAC基因在染色体上不均匀分布,MsNAC基因存在片段重复,并与大豆(Glycine max(Linn.)Merr.)、蒺藜苜蓿(Medicago truncatula)存在共线性关系,基因保守基序及基因结构分析表明MsNAC基因多数含有Motif1,Motif3保守基序及3~6个外显子。大多数MsNAC蛋白定位于细胞核中。系统进化树分析结果表明MsNACs聚类为8个亚家族。顺式作用元件分析表明MsNAC基因含有响应干旱及脱落酸、生长素等顺式作用元件。荧光定量分析结果显示部分MsNAC基因响应干旱、盐胁迫及脱落酸处理,大部分MsNAC候选基因在干旱、盐及ABA处理后上调表达。本研究结果可为紫花苜蓿抗旱、耐盐分子育种提供候选基因。

观赏植物NAC转录因子的研究进展

NAC转录因子家族在调节植物的生长发育中发挥着重要的作用,在模式植物、作物中已进行了大量研究,但是在观赏植物中的研究还缺乏系统性的梳理和探讨。本综述介绍了NAC转录因子的结构和分类,并梳理了2004-2023年NAC转录因子在观赏植物器官生长发育和胁迫响应上的生物学功能研究,其中观赏植物器官生长发育主要集中在叶缘形态建成、花器官发育、叶片衰老、花瓣衰老、种球休眠5个方面,胁迫响应则集中在干旱、盐、碱、冷、热等非生物胁迫,在生物胁迫中报道较少。最后,鉴于观赏植物NAC转录因子大部分都还停留在生物信息学分析以及表达模式分析等功能初探阶段,本文在结合观赏植物全基因组测序继续开展NAC转录因子鉴定研究、挖掘观赏植物中与模式植物存在不同作用机制的NAC转录因子、解析观赏植物NAC转录因子与其他转录因子间的调控网络、加快利用推进基因工程或编辑技术开展观赏植物的分子育种工作等4个方面,对未来NAC转录因子在观赏植物中的研究提出了展望。

植物NAC转录因子的研究进展

NAC转录因子家族是大型植物特异性转录因子家族之一,广泛参与植物侧根生长、次生壁形成、叶片衰老、花器官形成、果实发育等生长发育过程,并响应干旱、寒冷、盐碱以及病原菌入侵等逆境胁迫,极大地帮助了植物在各种环境中更好的生存。综述了近年来植物NAC转录因子家族结构和功能的研究进展,以期促进对NAC转录因子调控网络的理解和更深入的研究。

TaNAC069转基因抗叶锈病小麦种质的创制与分析

转录因子是植物中重要的一类抗病相关蛋白,其中NAC(NAM、ATAF1/2和CUC)类转录因子在植物应对生物胁迫和非生物胁迫方面起着重要作用。本研究前期获得了1个小麦(Triticum aestivum L.)NAC类转录因子TaNAC069,并初步证明其参与小麦抗叶锈病防御反应。本研究将TaNAC069基因构建到过表达载体pLGY-02中,利用农杆菌(Agrobacterium)介导的遗传转化法将pLGY-02-TaNAC069重组载体导入小麦感叶锈病农家品种‘JW’中,获得11个TaNAC069转基因T0株系。对T_(1)~T_(4)代植株进行PCR检测,明确TaNAC069已转入11个小麦株系中,并能稳定遗传;实时荧光定量PCR(Quantitative real-time PCR,qPCR)检测明确T1代11个转基因株系中TaNAC069基因的表达量显著高于受体材料‘JW’;对TaNAC069转基因小麦进行叶锈病抗性鉴定,TaNAC069转基因小麦叶锈病的发病程度比‘JW’轻,孢子量较少,表明TaNAC069基因过表达提高了转基因小麦对叶锈病的抗性。本研究明确了TaNAC069基因在小麦防御叶锈病过程中起正向调控作用,为解析TaNAC069的抗叶锈病机制奠定了基础。

茶树NAC转录因子基因CsNAC79和CsNAC9鉴定及其对非生物胁迫的响应

【目的】鉴定茶树NAC转录因子基因CsNAC79和CsNAC9,并分析它们在干旱、盐、高温、低温、外施ABA和外施GA3胁迫下的表达模式,为进一步解析CsNAC79和CsNAC9转录因子的生物学功能提供参考。【方法】以茶树‘龙井43’为材料,用RT-PCR扩增NAC转录因子家族基因CsNAC79和CsNAC9,并进行生物信息学分析;用qRT-PCR检测CsNAC79和CsNAC9在茶树6种非生物胁迫下的相对表达量。【结果】(1)CsNAC79和CsNAC9分别编码409个和282个氨基酸,且分别在15—150氨基酸位点、11—134氨基酸位点含有NAC家族成员典型的NAM保守结构域。(2)CsNAC79蛋白与中华猕猴桃、柿、洋蓟亲缘关系较近,CsNAC9蛋白与桃、荔枝、榴莲亲缘关系较近;2个NAC转录因子均是亲水性蛋白,皆不含信号肽和跨膜结构;CsNAC79和CsNAC9的启动子区域都含有非生物胁迫响应元件;蛋白质空间结构分析显示,CsNAC79和CsNAC9蛋白主要是由不规则卷曲和α-螺旋组成。(3)表达分析表明:CsNAC79和CsNAC9基因在6种不同非生物胁迫下均上调表达。【结论】茶树CsNAC79和CsNAC9基因响应多种非生物胁迫。

Transcriptome Profiling of Abscisic Acid-Related Pathways in SNAC4/9-Silenced Tomato Fruits

The NAC(NAM,ATAF,and CUC)family is considered one of the largest families of plant transcription factor,and its members are involved in fruit ripening.Abscisic acid(ABA)has been demonstrated to modulate the fruit ripening process.By applying the virus-induced gene silencing method and next-generation sequencing technology,we conducted a compara-tive analysis of the eff ects of SNAC4(SlNAC48,accession number:NM 001279348.2)and SNAC9(SlNAC19,accession number:XM 004236996.2)on tomato fruit ripening.The results of high-throughput sequencing identified 1262 significant(p<0.05)diff erentially expressed genes(DEGs)in SNAC4-silenced fruit compared to control fruit,while 655 DEGs were identified in SNAC9-silenced fruit.In addition,we selected 26 and 30 significant DEGs(p<0.05 and log 2-fold change>1.0)related to ABA in SNAC4-silenced and SNAC9-silenced tomatoes,respectively,for further analysis.The XET gene and two other genes(E8 and EXP1)were significantly down and upregulated in SNAC4-silenced tomatoes,respectively.However,the PYL9 gene and four other genes(PP2C,CYP707A2,EXPA6,and ACS6)were significantly down and upregulated in SNAC9-silenced tomatoes,respectively.In addition,ten DEGs were selected for use in tests to confirm the accuracy of the transcriptomic results by quantitative real-time polymerase chain reaction(qRT-PCR).Our results highlight the relationship between SNAC4/9 and ABA in the regulation of tomato ripening,which may help provide a theoretical basis for further research on the mechanisms of tomato fruit ripening and senescence.

NAC转录因子在水稻遗传改良中的研究进展

NAC转录因子是植物最大的特异性转录因子,具有特殊的NAC结构域和C端结构域,它们在调节水稻生长发育过程中发挥着重要的作用。目前在水稻中已鉴定到多种NAC转录因子,发现它们广泛参与调控水稻的产量及其品质性状,在水稻的遗传改良中具有重要功能。综述了NAC转录因子的分类与定位、调控水稻产量和稻米品质以及参与水稻非生物胁迫应答反应等新进展,以期为优质高产多抗水稻新品种的分子设计育种提供新的基因资源。

The NAC056 transcription factor confers freezing tolerance by positively regulating expression of CBFs and NIA1 in Arabidopsis

Freezing stress can seriously affect plant growth and development,but the mechanisms of these effects and plant responses to freezing stress require further exploration.Here,we identified a NAM,ATAF1/2,and CUC2(NAC)-family transcription factor(TF),NAC056,that can promote freezing tolerance in Arabidopsis.NAC056 mRNA levels are strongly induced by freezing stress in roots,and the nac056 mutant exhibits compromised freezing tolerance.NAC056 acts positively in response to freezing by directly promoting key C-repeat-binding factor(CBF)pathway genes.Interestingly,we found that CBF1 regulates nitrate assimilation by regulating the nitrate reductase gene NIA1 in plants;therefore,NAC056–CBF1–NIA1 form a regulatory module for the assimilation of nitrate and the growth of roots under freezing stress.In addition,35S::NAC056 transgenic plants show enhanced freezing tolerance,which is partially reversed in the cbfs triple mutant.Thus,NAC056 confers freezing tolerance through the CBF pathway,mediating plant responses to balance growth and freezing stress tolerance.

A NAC transcription factor MNAC3-centered regulatory network negatively modulates rice immunity against blast disease

NAC transcription factors(TFs)are pivotal in plant immunity against diverse pathogens.Here,we report the functional and regulatory network of MNAC3,a novel NAC TF,in rice immunity.MNAC3,a transcriptional activator,negatively modulates rice immunity against blast and bacterial leaf blight diseases and pathogen-associated molecular pattern(PAMP)-triggered immune responses.MNAC3 binds to a CACG cis-element and activates the transcription of immune-negative target genes OsINO80,OsJAZ10,and OsJAZ11.The negative function of MNAC3 in rice immunity depends on its transcription of downstream genes such as OsINO80 and OsJAZ10.MNAC3 interacts with immunity-related OsPP2C41(a protein phosphatase),ONAC066(a NAC TF),and OsDjA6(a DnaJ chaperone).ONAC066 and OsPP2C41 attenuate MNAC3 transcriptional activity,while OsDjA6 promotes it.Phosphorylation of MNAC3 at S163 is critical for its negative functions in rice immunity.OsPP2C41,which plays positive roles in rice blast resistance and chitin-triggered immune responses,dephosphorylates MNAC3,suppressing its transcriptional activity on the target genes OsINO80,OsJAZ10,and OsJAZ11 and promoting the translocation of MNAC3 from nucleus to cytoplasm.These results establish a MNAC3-centered regulatory network in which OsPP2C41 dephosphorylates MNAC3,attenuating its transcriptional activity on downstream immune-negative target genes in rice.Together,these findings deepen our understanding of molecular mechanisms in rice immunity and offer a novel strategy for genetic improvement of rice disease resistance.

植物NAC转录因子的生物学功能研究进展

转录因子(Transcription Factor,TF)是一类特殊结构的蛋白质分子,能够与特定DNA序列结合调控目标基因的特定表达.作为一类植物中特有的转录因子,NAC转录因子家族以保守的N端结构域和高度可变的C端转录激活域为特征,在调控植物的生长发育、生物和非生物胁迫响应以及次生代谢产物合成等过程中均发挥着至关重要的作用.文章总结前人研究,就NAC转录因子的结构特征及生物学功能的相关研究进展进行综述,以期深入探索NAC转录因子的调控机理,更好地了解其如何影响植物的生长发育和应对环境压力.

高州油茶NAC转录因子家族鉴定及生物信息学分析

NAC转录因子是植物中特有的转录因子,其在N端含有一段高度保守的特异结构域,对植物生长发育、胁迫反应和次生代谢产物生物合成等过程具有重要功能。研究基于高州油茶第三代全长转录组数据信息,通过生物信息学的方法筛选获得51个NAC转录因子,为深入研究基因家族特征,对其进行理化性质分析、多列比对分析、蛋白系统进化树、保守基序和基因结构分析、亚细胞定位,以及二级蛋白域结构预测、蛋白磷酸化位点分析。结果表明:NAC蛋白存在氨基酸数量差异,且氨基酸序列间疏水性呈现为亲水性蛋白,其蛋白结构中具有5个保守的亚结构域。NAC转录因子系统进化树分析表明116个基因可划分为10个亚族群,部分高州油茶NAC蛋白具有特异性聚为单一亚族群,其他与拟南芥和水稻融合为不同亚族群。同时NAC蛋白均具有NAM超家族结构。亚细胞定位表明51个高州油茶NAC蛋白均位于细胞核,且二级结构中随机卷曲结构为主,同时含有α-螺旋和β-转角结构等。蛋白磷酸化位点都具有丝氨酸(Ser)、苏氨酸(Thr)和酪氨酸(Tyr)。

Site-1 protease cleavage site is important for the ER stress-induced activation of membrane-associated transcription factor bZIP28 in Arabidopsis

Many sources of stress cause accumulation of unfolded or misfolded proteins in endoplasmic reticulum(ER), which elicits the unfolded protein response(UPR) to either promote cell survival or programmed cell death depending on different developmental context or stress severity. The Arabidopsis membrane-associated transcription factor, b ZIP28, is the functional equivalent of mammalian ATF6, which relocates from the ER to the Golgi where it is proteolytically processed and released from the membrane to the nucleus to mediate the UPR. Although the canonical site-1 protease(S1P) cleavage site on the ER lumen-facing domain is well conserved between b ZIP28 and ATF6, the importance of S1 P cleavage on b ZIP28 has not been experimentally demonstrated. Here we provide genetic evidence that the RRIL573 site, but not the RVLM373 site, on the lumen-facing domain of bZ IP28 is critical for the biological function of b ZIP28 under ER stress condition. Further biochemistry and cell biology studies demonstrated that the RRIL573 site, but not the RVLM373 site, is required for proteolytic processing and nuclear relocation of b ZIP28 in response to ER stress. Our results reveal that S1 P cleavage site plays a pivotal role in activation and function of b ZIP28 during UPR in plants.

葡萄NAC转录因子家族生物信息学分析

NAC基因家族是植物所特有的一类重要转录因子,广泛参与植物生长、发育以及器官建成、逆境胁迫应答等反应。目前,基因组信息的挖掘和分析已经成为葡萄功能基因组学研究的重要组成部分。本文利用生物信息学方法对葡萄143条NAC蛋白序列的系统发生和NAC基因组定位进行分析,然后对其氨基酸组成成分、理化性质以及二级和三级结构进行预测和分析,同时还分析了葡萄与拟南芥的NAC基因家族之间的联系。结果显示这143条蛋白序列与拟南芥94个NAC基因蛋白序列一起分成了16个亚族,说明拟南芥与葡萄NAC基因间具有较高的保守性。基因组定位结果发现143条NAC基因分布在17条染色体上。研究还发现不同亚家族间氨基酸数目、氨基酸序列间的疏水性存在一定的差异;而它们的二级结构都主要以随机卷曲为组成部分,且143条氨基酸序列三维结构十分相似。本文试验结果将为葡萄NAC基因家族的进一步功能分析提供重要研究基础。

植物NAC转录因子的结构功能及其表达调控研究进展

NAC转录因子是近十年来新发现的具有多种生物功能的植物特异转录因子.该家族转录因子的共同特点是其N端为保守的大约150个氨基酸的NAC结构域,C端为高度变异的转录调控区.它们在植物生长发育、激素调节和抵抗逆境等方面发挥着重要的作用.本文主要就植物NAC转录因子的基本结构特征、生物学功能、表达调控及其最新研究进展进行综述.

NAC转录因子在植物抗病和抗非生物胁迫反应中的作用

NAC转录因子是植物特有的一类转录因子,其共同特点是在N端含有一段高度保守、由约150个氨基酸组成的NAC结构域,而C端为高度变异的转录调控区。研究表明,NAC转录因子不仅参与植物生长发育的调控,而且在植物抗逆反应中具有重要的调控作用。文章着重介绍NAC转录因子在植物抗逆反应中的作用及其调控机制,并简要讨论NAC转录因子生物学功能的研究方向。

一个新的玉米NAC类基因(ZmNAC1)的克隆与分析

NAC转录因子是近十年来新发现的具有多种生物功能的植物特异转录因子,在植物生长发育、激素调节和抵抗逆境等方面发挥着重要的作用。本研究利用电子克隆方法,结合RT-PCR分析,从玉米中克隆了一个与NAC类基因同源的cDNA序列,命名为ZmNAC1(GenBank登录号:EU224278),该序列长度为1029bp,具有单一的完整开放阅读框架(ORF,26～907bp),推测编码蛋白含有293个氨基酸,分子量为32.3kDa,等电点为8.65。半定量RT-PCR分析表明,ZmNAC1可以被低温、聚乙二醇(Polyethylene glycol,PEG)、高盐和ABA诱导。这些结果表明,ZmNAC1可能参与植物逆境反应。本研究是有关玉米NAC转录因子基因的首次报道。