水稻HTD1基因启动子在转基因拟南芥中的表达特征

通过分析β-葡萄糖醛酸苷酶(β-glucuronidase, GUS）基因产物,对水稻HIGH-TILLERING DWARF1(HTD1)基因启动子在转基因拟南芥中的表达特性进行初步研究。用已构建的含HTD1基因启动子和GUS报告基因的植物表达载体,通过农杆菌介导转化拟南芥,对转基因拟南芥进行GUS组织化学染色,观察该启动子的表达特性。结果表明,在HTD1基因启动子的驱动下,GUS报告基因主要在转基因拟南芥幼苗期的叶片、叶柄、下胚轴以及主根基部的维管组织中表达。     

Construction of artificial promoters highly responsive to iron deficiency

Iron deficiency-responsive element 1 (IDE1) and IDE2 are cis-acting elements that are responsible for Fe-deficiency-inducible and root-specific expression of the barley (Hordeum vulgare L.) gene IDS2 (Fe-deficiency-specific clone no. 2). Using these cis-acting elements, we aimed to construct super-promoters that would induce prominent gene expression in the roots of Fe-deficient rice plants (Oryza sativa L.). Modules containing IDE1 and IDE2 of the IDS2 promoter were used as repeats or were linked to the Fe-deficiency-responsive promoter of barley IDS3, and were connected to known enhancer-like sequences. Five artificial promoters, as well as the native promoters of barley IDS2 or IDS3, were connected individually upstream of β-glucuronidase (GUS) and were introduced into rice. Transgenic rice plants were grown under control or Fe-deficient conditions, and GUS expression was analyzed. The artificial promoter that contained one module of IDE1 and IDE2 conferred strong Fe-deficiency-inducible GUS expression to the roots of rice plants. Each of the five artificial promoters induced a similar level of GUS expression in Fe-deficient roots, which did not exceed the GUS expression driven by the native IDS2 or IDS3 promoter. Artificial and native promoters induced GUS expression in response to Fe-deficiency in leaves, although the level of expression was lower than that in roots. Histochemical observations revealed that GUS expression driven by artificial and native promoters was spatially similar, and expression was dominant within vascular bundles and root exodermis. These findings suggest that there is coordinated expression of the genes that are involved in Fe-deficiency-induced Fe uptake in rice.     

Comparison of the functions of the barley nicotianamine synthase gene HvNAS1 and rice nicotianamine synthase gene OsNAS1 promoters in response to iron deficiency in transgenic tobacco

Barley ( Hordeum vulgare L.) nicotianamine synthase gene ( HvNAS1 ) expression in barley is strongly induced by Fe deficiency in the roots and rice ( Oryza sativa L.) nicotianamine synthase gene ( OsNAS1 ) expression in rice is induced by Fe deficiency both in the roots and in the shoots. In dicots, NAS genes are not strongly induced by Fe deficiency, and they function to maintain Fe homeostasis. Rice OsNAS1promoter::GUS or barley HvNAS1promoter::GUS was introduced into tobacco ( Nicotiana tabacum L.) and tissue specificities and systemic regulation of their expression were compared. A split-root experiment revealed that the HvNAS1 promoter exhibited functions similar to those of Fe-acquisition-related genes in tobacco roots, suggesting that this promoter responded to certain Fe-deficiency systemic signals and to the Fe concentration in the rhizosphere. The HvNAS1 promoter might harbor a type of universal system of gene expression for Fe acquisition. However, the OsNAS1 promoter did not respond to local application of Fe to the roots and induced GUS activities in mature leaves in response to Fe deficiency. This promoter might possess numerous types of cis -acting sequences that are involved in Fe metabolism.     

A rice FRD3-like (OsFRDL1) gene is expressed in the cells involved in long-distance transport

We identified four putative AtFRD3-like genes (OsFRDL) in the rice genome that exhibited 39.1 to 56.7% amino acid sequence similarities to Arabidopsis FRD3. Of these, we cloned three OsFRDL genes from a cDNA library prepared from iron-deficient rice roots: OsFRDL1, OsFRDL2, and OsFRDL3. OsFRDL1 was expressed weakly in Fe-sufficient roots, and slight expression was induced in the roots of Fe-deficient plants. OsFRDL2 was expressed constitutively in both roots and leaves, and Fe deficiency reduced its expression in leaves. OsFRDL3 was expressed in leaves, but not in roots; Fe deficiency induced slight expression in leaves. An OsFRDL1-sGFP fusion protein was localized in the plasma membrane in onion epidermal cells. The promoter GUS analysis showed that OsFRDL1 was localized in the cells involved in long-distance transport, in both Fe-sufficient and Fe-deficient plants. Furthermore, OsFRDL1 expression was observed during the reproductive stage. These results suggest that OsFRDL1 is a transporter that resides in the plasma membrane of cells involved in long-distant transport.     

粤油7号花生AhNCED1基因启动子克隆及其活性分析

AhNCED1是干旱胁迫下花生中调控脱落酸（abscisic acid,ABA）生物合成的关键基因。本文采用基于PCR的基因组DNA步移法,从抗旱性强的粤油7号花生中克隆AhNCED1基因起始密码子ATG上游2392bp启动子序列,构建该启动子驱动报告基因GUS的植物双元表达载体pAhNCED1p∷GUS,转化野生型拟...     

向日葵种子特异性启动子Ha ds10G1的克隆及其功能验证

利用PCR技术从向日葵基因组DNA中克隆了Lea蛋白基因家族中Ha ds10 G1基因上游1414bp的调控序列，序列分析表明-1—1414bp与报道序列同源性为100%，将其与GUS基因融合构建植物表达载体后，通过农杆菌介导法转化烟草NC89，PCR扩增初步证明目的片段已整合到烟草基因组中，转基因植株的GUS活性检测表明，在茎、叶中无GUS活性，GUS活性只存在于种子中，因此，Ha ds10 G1启动子具有种子特异性的功能。     

花生种子特异启动子AHSSP1的克隆及功能分析

为丰富花生种子特异启动子资源,本研究利用PCR技术在花生基因组中克隆了种子贮藏蛋白基因PSC32的启动子AHSSP1,利用半定量RT-PCR检测了PSC32基因表达模式,借助NewPLACE在线分析了AHSSP1序列中存在的顺式作用元件,并构建了AHSSP1驱动GUS报告基因的表达载体,经农杆菌转化获得转基因拟南芥,经GUS组织化学染色鉴定了该启动子的功能。结果表明,PSC32基因957 bp长的启动子AHSSP1序列具备种子特异表达启动子特有的3个RY REPEAT元件。半定量RT-PCR分析发现,PSC32基因在花生成熟种子中表达,而在饱果成熟期根、茎、叶片、花、入土前的果针、成熟种子的果壳中均不表达。GUS组织化学染色发现,转基因拟南芥成熟种子以及萌发种子的子叶、下胚轴和胚根均能够被染上蓝色;长出真叶后,子叶和下胚轴仍能被染色,而根和真叶不能被染上蓝色;成年期转基因拟南芥的叶片也不能被染上蓝色。而野生型拟南芥整个生长时期均不能被染上蓝色。以上现象说明AHSSP1是一个种子特异启动子。本研究丰富了花生种子特异启动子的资源,对花生籽仁品质改良或以花生籽仁作为“生物反应器”的研究具有重要的应用价值。     

Silicon alleviates long-term copper toxicity and influences gene expression in Nicotiana tabacum

Silicon (Si) is beneficial for plant growth and aids in stress tolerance. In this study, the effects of Si on long-term copper (Cu) toxicity in the low Si accumulator Nicotiana tabacum were evaluated. Silicon supplementation alleviated growth inhibition in roots and shoots of N. tabacum exposed to Cu toxicity. Alleviation of Cu toxicity correlated with increased Si accumulation in roots and leaves, suggesting N. tabacum contains a stress-regulated mechanism for Si transport. Root Cu concentration decreased in Si-supplemented plants exposed to Cu toxicity. Interestingly, Copper Transporter 1 (COPT1) expression decreased in roots of Si-supplemented plants exposed to Cu toxicity, which may contribute to Cu uptake reduction. Decreases in ethylene (ET) biosynthetic gene expression were previously implicated in Si-mediated stress alleviation. In the present study, Si-mediated alleviation of Cu toxicity corresponded with increased ET biosynthetic gene expression.     

Localization of MT2a Gene promoter expression is different from the site(s) of copper accumulation in the roots of transgenic Arabidopsis thaliana

Accumulation of mRNA for MT2a, a metallothionein of Arabidopsis thaliana, is known to be upregulated by excess copper (Cu). We compared the localization of the promoter activity of the MT2a gene and the accumulation site(s) of CU in the roots of transgenic A. thaliana. Open reading frame of the β-glucuronidase (GUS) gene was fused with the MT2a gene promoter and introduced into A. thaliana. Strong GUS activity was observed in the region near or within the stele in the presence of CU at a high concentration (high-CU condition). Electron probe X-ray micro-analysis demonstrated that under high-CU conditions, CU accumulated most abundantly in the cortex, where GUS activity was not significantly induced. These results indicate that the sites showing the MT2a promoter activity and CU accumulation did not coincide with each other, suggesting that CU is not a direct activator of the MT2a promoter.     

水稻OsWRKY19基因启动子的分析

利用PCR技术从水稻(Oryza sativa)秀水11品种中克隆了转录因子WRKY19编码区5'上游大小为1 404 bp的调控序列，命名为OsW19p, 将它和长度为105、378、977、1 106、1 205和1 306 bp的5'端缺失体分别与gus基因融合，构建植物表达载体。用根癌农杆菌介导法将所有载体转化水稻，GUS组织化学分析和荧光测定结果表明：(1)培养基中的2,4-D强烈抑制 gus基因在愈伤组织阶段的表达；(2) gus基因在转基因植株的根、茎、叶和花中均有表达，在花粉和成熟种子中无表达，在种子萌发时胚有表达，在苗期种子根和不定根及它们的侧根均有表达但根尖无表达，成熟期根部只有侧根有表达；(3)除p105以外OsW19p (p1404)和其它5个不同长度的缺失体均可驱动gus基因在转基因苗叶片中的表达，p1205活性最高，p977和p1306的活性减弱,由此推测-1404～-1306 bp和-1205～-977 bp间包含正调控元件，-1306～-1205 bp和-977～-378 bp间包含负调控元件。     

PMI基因为选择标记植物表达载体的构建及在雪柑转基因中的应用

以PMI基因替换植物表达载体pCAMBIA1301中的hpt基因以及pBI121中的GUS基因，构建了以PMI基因为选择标记基因的植物表达载体pCAMBIA1301PMI和pBIPMI，并导入根癌农杆菌EHA105中。研究了两种表达载体对雪柑上胚轴的转化，在培养基附加25 g•L-1甘露糖和5 g•L-1蔗糖为碳源的选择压下，pCAMBIA1301PMI的转化率为27.7％，pBIPMI的转化率为12.7％，对再生植株进行了氯酚红检测和PCR检测，建立了以PMI/甘露糖为选择系统的雪柑转基因体系。     

内含子数量改变GUS基因的瞬时表达调控

内含子是基因的重要组成部分,它与功能基因表达之间的关系越来越被重视。本研究以pCAM-BIA3301为载体,利用玉米泛素启动子Ubi1和水稻肌动蛋白启动子Actin1构建两个GUS基因表达载体p33U1和p33A1,同时设置3个对照载体。通过基因枪轰击法将上述载体转入水稻胚性愈伤组织,探讨内含子对外源目的基因表达的调控作用。组织化学检测结果表明：CaMV35S启动子调控下的iGUS（带内含子）基因能够顺利表达;同样,Actin1启动子（带内含子）调控下的不带内含子的GUS基因也可以正常表达,而当Actin1启动子（带内含子）驱动iGUS基因时,则导致GUS染色反应不能发生。Ubi1启动子（带内含子）调控GUS基因的瞬时表达也得出类似结果,证明表达框中内含子的数量为1个或两个时,对GUS基因的表达起到了不同的调控作用。本研究结果对植物表达载体构建及功能基因表达都具有指导意义。     

冷诱导基因转录因子CBF1的组成型表达对植物的抗寒性及生长发育的影响

通过农杆菌介导法将玉米泛素启动子(Pubi)调控的CBF1基因以及CaMV35启动子调控的GUS基因一起转化烟草,研究了冷诱导基因的转录因子CBF1(CRT/DRE binding factor 1)对植物抗寒性及生长发育的影响。结果表明:CBF1的组成型表达增强了冷敏感植物的抗寒性;在表型上使植株矮壮,叶片着生密集,叶色深绿,叶厚度增加;促进侧枝生长,延长营养生长期,推迟花期;对植株的结实性无明显影响。植株抗寒能力以及表型变化程度与CBF1的表达强度相关。因此,利用CBF1基因进行植物遗传改良时应根据物种的应用特点采用合适的表达调控策略。     

外源NO对铜胁迫下小白菜SBPase基因表达特性及其光合速率的影响

【目的】本研究通过克隆小白菜光合暗反应中限制核酮糖-1, 5-二磷酸(RuBP)再生的关键酶景天庚酮糖-1,7-二磷酸酶(sedoheptulose-1, 7-bisphosphatase,SBPase)基因,分析铜胁迫及添加外源一氧化氮(NO)供体硝普钠(SNP)缓解铜胁迫时该基因的表达情况,并将其与对应处理下小白菜净光合速率(Pn)的变化情况结合在一起进行分析,以期为全面了解外源NO缓解铜胁迫植物光合作用机理提供理论依据。【方法】以小白菜品种“上海青”3~4片真叶大的幼苗为材料,采用RT-PCR技术克隆小白菜SBPase基因,铜处理浓度为200 μmol/L,外源NO供体SNP浓度为300 μmol/L,同时设置相关的4个对照组排除其他可能因素的干扰,在添加处理液后的0、 4、 8、 12 d上午九点测定各处理小白菜相同节位叶片的净光合速率,并利用实时荧光定量PCR技术检测在对应的处理时间及对应节位小白菜叶片中SBPase基因的表达情况。试验环境条件为光照强度200 μmol/(m2·s),光周期12 h,温度25℃/18℃。【结果】 1)克隆得到小白菜SBPase基因(Genbank登录号:AHY18974.1),开放阅读框为1182 bp,编码393个氨基酸,蛋白质分子量为42.34kD,理论等电点为5.85。2)序列分析表明其含有氧化还原活性位点,包含一个具有59个氨基酸残基的叶绿体转移肽序列。小白菜SBPase基因推导的氨基酸序列与其他物种中已分离的SBPase编码的氨基酸序列具有很高的同源性。3)实时荧光定量PCR分析表明,SBPase基因在铜胁迫下的小白菜叶片中表达量下降,且随着胁迫时间的延长下降程度加剧;而在铜胁迫的基础上添加外源NO可以在一定程度上缓解铜胁迫引起的小白菜叶中SBPase基因表达量的下降。4)铜胁迫下小白菜叶片Pn显著下降,且随着胁迫时间的延长,Pn下降加剧,施加外源NO后Pn的下降得到一定程度的缓解,各处理下Pn的变化与SBPase基因表达量变化趋势一致。【结论】铜胁迫及在铜胁迫的基础上添加外源NO后小白菜叶片中SBPase基因表达量的变化,可能是影响对应条件下小白菜叶片的净光合速率变化的原因之一。     

Seasonal variation of defense-related gene expression in leaves from Bois noir affected and recovered grapevines

Although Bois noir is one of the main phytoplasma diseases of grapevine, the gene expression and enzyme activities that underlie physiological changes occurring in symptomatic and recovered (with spontaneous or induced symptom remission) plants are mostly unknown. Bois noir symptomatic leaves (September 2006, 2007) and symptomless leaves from infected symptomatic plants (September 2007) of Sangiovese (moderately susceptible) and Chardonnay (highly susceptible) cultivars were collected. Moreover, leaves from infected symptomless plants of both cultivars were harvested in June 2007. Leaves from recovered plants were also collected in the same periods. In recovered plants of both cultivars, class III chitinase and almost every time phenylalanine ammonia-lyase and chalcone synthase expression were increased for all collection periods. In symptomatic leaves of both cultivars, the expressions of the same genes were up-regulated and also those of β-1,3-glucanase and flavanone 3-hydroxylase. The activities of chitinase, phenylalanine ammonia-lyase, β-1,3-glucanase, and superoxide dismutase generally correlated with gene expression. For the moderately susceptible Sangiovese, the defense genes were generally up-regulated in both symptomatic and symptomless leaves (for all collection periods). This behavior was not observed in the highly susceptible Chardonnay, in which changes in gene expression were linked to evident symptom display. Therefore, the physiological response of the plants to this pathogen infection appear to be the reason for the resistance of the cultivar to the disease.     

Dynamics of nitrogen translocation from mature leaves to new shoots and related gene expression during spring shoots development in tea plants (Camellia sinensis L.)

The contribution of N remobilization is crucial for new shoots growth and quality formation during spring tea shoots development. However, the translocation mechanism of N from source leaves to sink young shoots is not well understood. In the present study, ¹⁵N urea was applied to mature tea leaves one week before bud break to track N remobilization in a field experiment. The dynamic changes in plant ¹⁵N abundance, contents of amino acids, and the expression levels of genes related to N metabolism and translocation were followed during the 18‐d development of new spring shoots until three expanding young leaves. The results showed that during the growth of new shoots the amount of ¹⁵N in the shoots increased, whereas the N_dff (N derived from ¹⁵N‐urea) in mature leaves decreased, showing that the foliar‐applied N in mature leaves was readily exported to new shoots. This process was found to be accompanied by decline of chlorophylls. In the mature leaves, expression CsATG18a and CsSAG12 involved in autophagy was dramatically induced (> 4‐fold) at approximately nine days after the bud breaking. The genes involved in the transformation of amino acids, including primarily CsGDH2, CsGDH4, CsGLT3, CsGS1;3, and CsASN2 were upregulated by > 3‐fold after bud breaking. The expression levels of CsATG8A, CsATG9, CsSAG12, CsGS1;1, CsGDH1, and CsAAP6 correlated negatively with the N_dff in mature leaves, but positively with ¹⁵N amount and total N amount in new shoots, suggesting these genes played important roles in N export from mature leaves. In the new shoots, the expression of most genes showed two defined peaks, one on six days and one on 12 days after bud breaking. The expression of CsGS2, CsASN3, CsGLT1, and CsAAP4 positively correlated with the ¹⁵N amount and total N amount in new shoots. These genes might be involved in the transport and re‐assimilation of N from mature leaves. The overall results demonstrated that the translocation of ¹⁵N from mature leaves to new spring shoots was regulated by the genes involved in autophagy, protein degradation, amino acid transformation and transport.     

杉木紫色酸性磷酸酶基因ClPAP18b的克隆及表达分析

  【目的】  以速生丰产型杉木无性系洋023、洋036、洋6421和拟南芥为材料,研究杉木中紫色酸性磷酸酶 (PAPs) 的功能作用,以筛选具有磷素高效利用特性的杉木无性系。  【方法】  采用PCR技术克隆PAP18b基因,分析其序列特征和同源性,并对杉木无性系洋023、洋036、洋6421进行正常供磷 (1.0 mmol/L KH₂PO₄) 和低磷胁迫 (0.1 mmol/L KH₂PO₄,0.9 mmol/L KCl) 砂培盆栽处理0、10、15、30和60天,测定酸性磷酸酶活性及全磷含量,定量分析根和叶中ClPAP18b基因表达量、磷含量及酸性磷酸酶活性的关系,并将ClPAP18b基因过表达至拟南芥,进行该基因的功能验证。  【结果】  成功克隆获得杉木PAP18b基因CDS序列 (1 212 bp),命名为ClPAP18b,该基因编码404个氨基酸,亚细胞定位于胞间区,这表明ClPAP18b基因可能发挥调控酸性磷酸酶分泌至胞外的功能。酸性磷酸酶活性测定结果表明,30天磷处理后,正常供磷和低磷处理下,杉木洋036和洋6421无性系根中的酸性磷酸酶活性均高于叶,而根中酸性磷酸酶活性低磷处理下高于正常供磷处理;洋023的根和叶中酸性磷酸酶活性在低磷胁迫诱导下多高于正常供磷条件下根和叶中酸性磷酸酶活性。磷含量分析结果表明,杉木洋023、洋036和洋6421无性系的地上部磷含量高于地下部,不同水平供磷处理后,不同杉木无性系或同一杉木不同组织磷含量存在差异。RT-qPCR 结果显示,低磷胁迫诱导杉木ClPAP18b基因表达。低磷条件下,ClPAP18b过表达拟南芥植株长势优于对照组,且过表达植株中的酸性磷酸酶活性叶高于对照组,但花青素积累量低于对照组。此外,相比于正常供磷处理植株,过表达ClPAP18b拟南芥中PHT1; 2、PHT1; 8和AtPAP26等与磷胁迫相关的基因表达量显著高于对照组,而AtPAP12和AtPAP17基因表达量明显降低。  【结论】  低磷胁迫诱导杉木ClPAP18b基因表达和酸性磷酸酶活性增强,但不同杉木无性系对磷缺乏的适应性存在明显差异,过表达ClPAP18b基因可促进拟南芥植株耐低磷胁迫,ClPAP18b基因可能在杉木低磷胁迫调节机制中发挥调控作用,可作为改良杉木耐低磷的重要候选基因。     

遗传转化KN1基因促进毛果杨外植体再生

已有研究表明玉米同源异形盒（homeobox）基因Knotted1（KN1）超量表达会导致转基因烟草、拟南芥和番茄等植物中细胞分裂素含量增加,提高转化效率。由于木本植物遗传转化困难,且毛果杨可作为木本植物研究的模式植物,因此,本研究利用农杆菌介导法,将35S启动子驱动的玉米KN1（35S：：KN1）基因导入毛果杨,并分析KN1基因表达对毛果杨再生率的影响。研究结果表明,经GUS组织化学染色及PCR分子鉴定,KN1基因已经成功导入毛果杨幼苗;KN1基因对毛果杨外植体愈伤诱导率影响不明显,但单个外植体芽诱导率比对照高0.96倍,毛果杨的再生频率明显提高;与移栽成活的野生对照植株相比,转化KN1基因植株出现叶片变小,叶色变深,在叶片边缘产生裂片,分枝增加,无顶端优势等特殊表型。本研究中KN1基因引起转化植株表型变化,因此在毛果杨的遗传转化中,可作为一种有效的正向选择标记基因。     

香蕉乙烯受体基因RNA干扰表达载体的构建

设汁两对引物，PCR扩增香蕉乙烯受体基因cDNA序列自AUG启始密码子起长538bp区段的正向(记为5‘I)、反向(记为AI’)DNA区段。构建含该正、反向互补重复序列DNA片段的中间载体，经测序证实连接正确后。克隆到植物表达载体pCAMBIA-1301中的GUs基因位置并经双酶切证实。在所得表达载体pCAMBIA-1301-S5’I-AI’中，该插入正反互补重复片段处于35S启动子之后，由此转录的mRNA因两端序列反向互补而形成发夹式RNA，因此可应用于RNA干扰研究。同时，文中对构建含正、反向重复序列插入片段植物表达载体过程中出现的插入片段链内退火引起连接困难等问题进行了分析讨论。     

烟叶成熟期氮代谢酶活性、基因表达与烤烟氮素利用效率的关系

【目的】 谷氨酰胺合成酶 (NtGS1和NtGS2) 和硝酸还原酶 (NtNit) 基因的表达丰度反映了作物氮素代谢的特征。研究不同氮效率烤烟品种成熟期叶片的氮代谢相关酶活性和相关基因的表达,加深理解不同烤烟品种叶片氮素代谢的生理生化机制,为耐氮肥品种选育提供理论基础。 【方法】 以氮效率不同的三个烤烟品种豫烟10号、K326和NC89为材料进行盆栽试验。在叶龄35、45、55、65、75 d (以幼叶长1 cm、宽0.5 cm时作为叶龄第1天) 取第12片叶 (自下向上数),采用实时荧光定量PCR技术,测定叶片谷氨酰胺合成酶基因 (NtGS1和NtGS2) 和硝酸还原酶基因 (NtNit) 的表达丰度,同时采用常规生理生化测试技术对叶片酶活性和NH₄+ 浓度、总氮、可溶性蛋白、质外体相关指标以及氨气挥发量进行测定。 【结果】 各品种NtGS1表达丰度自叶龄35 d始大幅上调,于55 d达到最大值后表达下调。NtGS2表达丰度自35 d始呈持续下调表达。NtNit表达整体变化趋势与NtGS2相似。在叶龄45～65 d,上述三种基因的表达丰度均表现为NC89 > K326 > 豫烟10号,且品种间表达差异达极显著水平。 NtGS1基因表达丰度与叶片总氮、叶片铵浓度、质外体铵浓度以及氨气挥发量呈极显著负相关,与NR活性和质外体pH呈显著负相关。NtGS2基因表达丰度与GS活性、总氮、可溶性蛋白含量和质外体pH呈极显著正相关,与NR活性、叶片铵浓度呈显著正相关,而与质外体铵浓度和氨气挥发量呈显著负相关。NR基因表达丰度与总氮、可溶性蛋白含量、叶片铵浓度呈显著正相关,与氨气挥发量呈显著负相关。 【结论】 氮低效烤烟品种成熟期叶片中两种谷氨酰胺合成酶同工酶活性均较低,氮素转移和再利用能力差,导致植株吸收的氮素以氨气形式挥发损失量大,叶片衰老速度较快。而氮高效品种氮素同化和再利用能力较强,氨气挥发量小,易发生贪青晚熟。