刚毛柽柳NAC24基因的表达及抗逆功能分析

【目的】NAC类转录因子是植物特有的最大的转录因子家族之一,广泛参与植物的生长发育过程,并在植物响应盐、干旱等多种非生物胁迫的过程中发挥至关重要的调控作用。本研究拟从盐生木本植物刚毛柽柳中克隆获得一个NAC转录因子基因,研究该基因的耐盐、抗旱功能,以期为研究木本植物NAC转录因子的抗逆分子机制奠定理论基础。【方法】在刚毛柽柳NaHCO_3胁迫转录组数据库中筛选获得一个NAC转录因子基因,将其命名为ThNAC24(GenBank登陆号:KF031949)。利用生物信息学工具将其与其他9个物种的NAC蛋白进行多序列比对,与拟南芥105个NAC蛋白进行进化树分析。分别用300 mmol·L^-1 NaCl和400 mmol·L^-1甘露醇对刚毛柽柳进行胁迫,在胁迫6、12、24和48 h后分别取刚毛柽柳根及叶组织。通过实时荧光定量RT-PCR(qRT-PCR)技术分析盐、干旱胁迫下ThNAC24基因在不同胁迫时间点及不同组织的表达情况,初步鉴定其是否响应盐、干旱胁迫。为进一步研究ThNAC24基因的抗逆功能,分别构建植物过表达(pROKⅡ-ThNAC24)及抑制表达(pFGC5941-ThNAC24)载体。利用农杆菌介导的高效瞬时遗传转化体系获得ThNAC24基因瞬时过表达(OE)、抑制表达(IE)及对照(Control)刚毛柽柳植株。在盐、干旱胁迫下分析比较了ThNAC24基因瞬时过表达、抑制表达及对照刚毛柽柳植株的二氨基联苯胺(DAB)和氯化硝基四氮唑蓝(NBT)染色情况,过氧化物酶(POD)和超氧化物歧化酶(SOD)活性,及电解质渗透率、失水率及丙二醛(MDA)含量,鉴定ThNAC24基因的耐盐、抗旱功能。【结果】ThNAC24基因的开放阅读框为1 023 bp,编码340个氨基酸。多序列比对结果显示ThNAC24在N端的氨基酸序列相似度比较高,具有NAC家族的序列特征;系统进化树分析结果显示ThNAC24与ANAC103和ANAC082的亲缘关系较近。qRT-PCR结果显示:盐胁迫下,ThNAC24基因上调表达,在根组织中胁迫12 h表达量最高,而叶组织中胁迫24 h的表达量最高;干旱胁迫下,ThNAC24基因上调表达,在根组织中胁迫6 h表达量最高,在叶组织中胁迫12 h的表达量最高。ThNAC24基因在刚毛柽柳根和叶组织中均有表达且响应盐和干旱胁迫。过表达ThNAC24基因显著降低了刚毛柽柳H_2O_2和超氧阴离子含量,增强了POD和SOD酶的活性,从而减少活性氧(ROS)的积累。过表达ThNAC24基因能够降低刚毛柽柳在逆境胁迫下的电解质渗透率、失水率及MDA的积累,从而保护细胞膜结构的完整性。【结论】刚毛柽柳ThNAC24基因能够响应盐、干旱胁迫,过表达ThNAC24基因植株通过增强POD和SOD活性,进而提高ROS清除能力,减少细胞受损或死亡,从而提高刚毛柽柳的耐盐及抗旱能力。     

胁迫条件下毛竹miR164b及其靶基因PeNAC1表达研究

MicroRNAs(miRNAs)在植物生长发育以及抗逆等过程中起着重要的调控作用。miR164 作为植物特有的miRNA,其主要的靶基因是植物NAC转录因子。为揭示毛竹 miR164 对其靶基因的调控机制,通过茎环引物法和RT-PCR技术,从毛竹(Phyllostachys edulis)中克隆出miR164b成熟序列及其靶基因PeNAC1。序列分析表明miR164b的靶点位于PeNAC1 编码区,RLM-5'RACE PCR产物测序结果证实切割位点位于靶点的第10-11位碱基之间。组织特异性表达分析表明,毛竹 miR164b和PeNAC1在根、茎、叶及鞘中均表达,其中miR164b在根中表达丰度最高,在茎中表达丰度最低;而PeNAC1的表达丰度恰好与miR164b 相反。实时定量PCR结果显示,NaCl(250 mmol·L^-1)、低温(4℃)和强光(1 500 μmol·m^-2·s^-1)处理后毛竹叶片中 miR164b 的表达均明显下调,GA₃(100 μmol·L^-1)处理后 miR164b表达量明显上调;而在同样处理条件下,PeNAC1的表达恰好呈现出与其完全相反的趋势。由此表明,miR164b对靶基因PeNAC1 具有表达调控作用,可能与毛竹响应非生物胁迫的抗逆过程密切相关,这为利用miRNA开展竹子抗逆分子育种提供了参考。     

平榛ChWRKY28基因克隆及表达模式分析

[目的]研究平榛ChWRKY28基因序列特征及其在不同非生物胁迫下的表达规律.[方法]以平榛为试材,采用RACE-PCR方法进行基因克隆;利用实时荧光定量PCR方法检测基因在不同组织及不同非生物胁迫下的表达模式.[结果]表明:克隆得到的WRKY基因,全长1 342 bp,基因内部包含1个长963 bp的完整开放阅读框,编码320个氨基酸残基,命名为ChWRKY28.构建的系统发育树表明:该序列与拟南芥AtWRKY28及杨树PtrWRKY93的关系最近,相似性分别为49%和60%.基因表达分析表明:ChWRKY28在雄花序、雌花芽及茎中均有表达,但在茎部(皮)中的表达量高于雄花序和雌花芽中的表达量,具有组织表达特异性;低温、干旱及盐胁迫均能诱导ChWRKY28基因的表达,但受诱导程度存在差异.亚细胞定位分析结果表明:ChWRKY28蛋白分布在细胞核内,是一个核蛋白.[结论]推测ChWRKY28基因可能参与植物响应非生物胁迫的信号转导过程.     

思茅松1-脱氧-D-木酮糖-5-磷酸合酶(DXS)基因的克隆及功能分析

1-脱氧-D-木酮糖-5-磷酸合酶(DXS)是甲基-D-赤藓醇-4-磷酸(MEP)途径中的第一个酶,也是限速酶。本文根据思茅松(Pinus kesiya var. langbianensis(A. Chev.) Gaussen)树皮转录组数据分析结果,获得思茅松DXS基因片段,然后根据获得的基因片段设计特异引物,运用RT-PCR和RACE技术从思茅松树皮中克隆得到完整的DXS基因(PkDXS1)。PkDXS1基因的cDNA全长序列2888 bp,含有1个2223 bp的开放阅读框(ORF),编码740个氨基酸,该基因推断的蛋白与赤松(Pinus densiflora Siebold & Zucc)DXS蛋白的相似性为99%,与欧洲云杉(Picea abies(L.) H. Karst.)DXS的相似性为97%;经氨基酸序列比对,推断思茅松DXS具有高等植物DXS酶特有的叶绿体转运肽,二磷酸硫胺结合位点和转酮醇酶结构域。半定量RT-PCR检测表明树皮的创伤促进DXS基因的表达。     

核桃JreIF1A基因启动子的鉴定及表达活性分析

真核生物翻译起始因子(eIF1)对逆境响应具有一定正响应;启动子能有效预测基因功能。以本研究鉴定获得核桃eIF1A基因(即JreIF1A)为其上游启动子,通过生物信息学、瞬时转化及GUS活性测定等不同技术,分析JreIF1A启动子的基本生物功能。结果显示,JreIF1A上游1 200bp启动子序列中包含众多与逆境响应相关的元件,病害响应相关的BIHD1OS、热胁迫响应相关的CCAATBOX1、Dof转录因子识别的DOFCOREZM、WRKY识别的WRKY71OS等。将JreIF1A启动子瞬时转入烟草测定GUS酶活性,发现JreIF1A启动子具有表达活性,且受干旱、盐、寒、热等胁迫诱导。表明JreIF1A启动子具有逆境响应表达活性,可能调控JreIF1A基因参与多重逆境响应。     

刚毛柽柳ThP5CR基因的克隆及抗逆功能分析

【目的】脯氨酸在植物对盐和干旱胁迫应答中起重要的渗透调节作用,增加植物体内脯氨酸含量可以提高植物抗逆性。吡咯啉-5-羧酸还原酶(P5CR)是合成脯氨酸的重要还原酶,本研究拟从抗旱耐盐植物刚毛柽柳中克隆获得1个ThP5CR基因,研究该基因的抗逆功能,为该基因用于林木基因工程育种奠定理论基础。【方法】以"Pyrroline 5 Carboxylate Reductase"作为关键词,对实验室前期构建的刚毛柽柳转录组序列进行比对查找获得ThP5CR基因c DNA序列,并通过RT-PCR和测序验证克隆获得的ThP5CR基因序列。利用生物信息学工具进行序列分析。利用Real time RT-PCR分析不同逆境处理下ThP5CR基因在刚毛柽柳根和叶中的表达情况。为了进一步分析ThP5CR基因的抗逆功能,构建了植物过表达载体pROKⅡ-ThP5CR,并进行刚毛柽柳瞬时转化,同时以pROKⅡ瞬时侵染刚毛柽柳作为对照。分析比较Na Cl和甘露醇胁迫后2种瞬时转化刚毛柽柳的脯氨酸、MDA、H2O2含量和二氨基联苯胺(DAB)、氯化硝基四氮唑蓝(NBT)及伊文思蓝(Evans blue)染色情况,以综合评定ThP5CR基因的抗逆功能。【结果】ThP5CR基因编码273个氨基酸,编码蛋白分子量为28.29 k Da,理论等电点为9.22。含有典型的NADP结构域和P5CR二聚体结构域。2种胁迫(Na Cl和PEG6000)和3种激素(ABA、GA3和JA)处理后,刚毛柽柳ThP5CR基因的表达都发生了不同程度的改变,且每种处理后至少有1个时间点发生了明显改变。此外,Na Cl胁迫、PEG6000胁迫和ABA激素处理对ThP5CR基因的表达产生的影响更为显著。对2种瞬时表达刚毛柽柳植株中ThP5CR基因的表达分析显示成功获得瞬时过表达转基因株系(标记为OX)。进一步的生理指标和生理染色结果显示,非胁迫条件下,过表达植株脯氨酸含量高于对照植株(标记为Con);150 mmol·L-1Na Cl和200 mmol·L-1甘露醇胁迫后,2种转基因株系脯氨酸含量差异则更显著;NBT、DAB染色和H2O2含量测定结果显示,OX植株的O-·2和H2O2含量明显低于对照;而Evans blue染色结果和MDA含量测定表明,与对照相比,过表达ThP5CR植株着色更浅、MDA含量更低。【结论】ThP5CR基因能对Na Cl、PEG胁迫和ABA等激素处理做出应答,可能参与了刚毛柽柳抗旱耐盐胁迫应答。在150 mmol·L-1Na Cl和200 mmol·L-1甘露醇胁迫下瞬时过表达ThP5CR植株可通过增加脯氨酸含量增强细胞内清除活性氧能力,使O-·2和H2O2的积累减少,从而减少细胞受损或细胞死亡,增强植物的抗逆性。本研究初步证实ThP5CR基因是一个逆境胁迫应答候选基因。     

长白山典型森林土壤黑碳含量及不同组分中的分布特征

[目的]通过实地采样对黑碳进行量化研究。[方法]利用相对密度分组方法探讨了长白山典型森林土壤黑碳含量及在不同有机碳组分中的分布特征。[结果]表明:黑碳(BC)含量在表层(A₁₁)、亚表层(A₁₂)分别为6.39~16.55 g·kg^-1、1.44~6.16 g·kg^-1,随土壤深度的增加而显著下降(p﹤0.01)。在A₁₁、A₁₂层中,轻组有机碳(LFOC)平均含量分别为66.66 g·kg^-1、6.65 g·kg^-1,轻组黑碳(LFBC)平均含量分别为5.63 g·kg^-1、1.21 g·kg^-1,同时,LFBC/LFOC在A₁₂层(10.02%~34.89%)显著高于A₁₁层(6.99%~14.45%)(p﹤0.01);A₁₁、A₁₂层重组有机碳(HFOC)平均含量分别为49.16 g·kg^-1、36.55 g·kg^-1,重组黑碳(HFBC)平均含量分别为2.69 g·kg^-1、1.44 g·kg^-1,HFBC/HFOC在A₁₁层(3.36%~8.08%)和A₁₂层(3.21%~7.58%)之间没有显著差异(p>0.05)。另外,土壤中LFBC/LFOC显著高于HFBC/HFOC(p﹤0.01),LFBC/BC显著高于HFBC/BC(p﹤0.01)。[结论]长白山典型森林土壤中黑碳的含量、比例均较高;土壤表层(A₁₁)有机碳、黑碳含量及各组分有机碳、黑碳含量均高于亚表层(A₁₂),均随着土层加深而显著降低;轻组、重组有机碳中均含有一定比例的黑碳,黑碳主要分布在轻组分中;轻组、重组有机碳与组分中黑碳均显著相关,轻组中相关系数大于重组。     

绿竹BoGPIAP基因克隆与异位表达研究

糖基化磷脂酰肌醇锚定蛋白(GPIAP)因其结构和功能的多样性,决定了它在各种生物学过程中都发挥着重要作用。采用同源克隆的方法从绿竹(Bambusa oldhamii)中获得一个GPIAP同源基因,命名为BoGPIAP,cDNA全长1 772 bp,其中包括1 356 bp的开放阅读框,编码一个451 aa的的蛋白。蛋白结构分析表明,该蛋白包含1个典型的GPIAP家族保守区域(47-211)和1个CCVS结构域,在N-端和C-端分别具有1个跨膜信号肽和1个GPI锚定信号肽,属于GPIAP家族。构建BoGPIAP与GFP融合的表达载体,在洋葱表皮细胞中瞬时表达,结果显示BoCOBL::GFP融合蛋白定位于细胞膜上,证明BoGPIAP基因编码的蛋白为膜蛋白。分别构建BoGPIAP的正义、反义表达载体并转化烟草(Nicotiana tabacum)。PCR检测结果表明,BoGPIAP已转入烟草。与野生型相比,转反义基因植株细弱,纤维细胞壁明显变薄;而转正义基因植株粗壮,纤维细胞壁明显变厚。表明BoGPIAP可能对竹子纤维细胞壁的发育具有调控作用。     

林木非生物胁迫抗性基因工程研究进展

干旱、极端温度和盐害等非生物胁迫因子是制约林木生长的重要因素。由于林木生长周期长, 且抗逆机制极为复杂, 长期以来, 如何改良林木对非生物胁迫的抗性一直是育种学家的难题。然而, 随着基因工程技术的发展, 人们可以在基因水平上改造林木, 提高其抗逆能力。文中主要介绍了林木抗逆基因工程的研究进展, 探讨了目前基因工程技术应用于林木抗逆育种研究存在的问题, 并对其应用前景进行了展望。     

An integrated model for predicting maximum net photosynthetic rate of cocksfoot (Dactylis glomerata) leaves in silvopastoral systems

Net light-saturated photosynthetic rate (A_max) of field grown cocksfoot (Dactylis glomerata L.) leaves in a radiata pine (Pinus radiata D. Don) silvopastoral system (Canterbury, New Zealand) was measured at different times under severe shade (85–95 μmol m^–2 s^–1 photosynthetic photon flux density, PPFD) and in full sunlight (1900 μmol m^–2 s^–1 PPFD). The aim was to integrate individual functions for A_max against air temperature (2 to 37 oC), water status, expressed as pre-dawn leaf water potential (ψ_lp) (-0.01 to −1.6 MPa), herbage nitrogen (N) (1.5 to 5.9%), regrowth duration (20 to 60 days) and time under shade (1 to 180 min) into a multiplicative model. The highest A_max value obtained was 27.4 μmol CO₂ m^–2 s^–1 in non-limiting conditions with full sunlight. This value was defined as standardised dimensionless A_maxs = 1 for comparison of factor effects. The canopy temperature of the cocksfoot sward was up to 7.4 oC cooler than air temperature for plants under shade. Therefore, canopy temperature was used to predict A_max. The only interaction was between time under severe shade (5% of the open PPFD) and water stress (ψ_lp = −0.4 to −1.3 MPa) and this was included in the model. Validation of this model indicated 78% of the variation in A_max could be accounted for using these five factors by the addition of the interaction function. This model could be used to assist the prediction of pasture growth in silvopastoral systems through incorporation into a canopy photosynthesis model. This revised version was published online in June 2006 with corrections to the Cover Date.     

麻疯树BRI1基因的鉴定及其在不同发育时期花蕾中的表达分析

[目的]研究油菜素内酯受体BRI1的同源基因(JcBRI1)在麻疯树花发育过程中的作用。[方法]利用RT-PCR技术克隆JcBRI1基因的CDS,以pET-30a(+)质粒为框架构建原核表达载体转化至大肠杆菌进行诱导表达,随后利用LC-MS/MS对表达产物进行质谱鉴定,并以氨基酸序列为基础分析JcBRI1蛋白质结构。利用荧光定量PCR分析麻疯树JcBRI1基因在雌雄花发育的关键时期的表达水平,初步确定其在麻疯树花发育过程中的表达模式。[结果]克隆获得了JcBRI1基因的CDS,长度为3 591 bp。SDS-PAGE检测结果表明:JcBRI1基因在大肠杆菌BL21(DE3)和Rosetta(DE3)中均能表达,但在低温环境的表达量更高。原核表达产物的LC-MS/MS鉴定结果表明:JcBRI1氨基酸序列与预期的一致,JcBRI1基因的表达产物为麻疯树BRI1蛋白。对JcBRI1在麻疯树花器官不同发育时期的表达水平进行检测分析,结果表明:JcBRI1基因的表达量在雌花发育的第一个时期即大孢子母细胞时期就达到了最高值,在随后几个时期持续下调;然而,其在雄花各个时期的表达量并无明显差异,表明其很有可能参与了麻疯树雌花大孢子母细胞发育过程。[结论]麻疯树JcBRI1基因为LRR-RLKs家族成员BRI1的同源基因,很有可能参与麻疯树雌花大孢子母细胞的发育过程,至于JcBRI1在麻疯树大孢子母细胞发育过程中的具体作用机制还需进一步研究。     

中间锦鸡儿CiDR1的克隆及干旱胁迫下的表达分析

[目的]研究并了解中间锦鸡儿CiDR1基因功能及其对干旱胁迫的响应,为抗性育种提供候选基因。[方法]通过RACE技术从中间锦鸡儿中克隆CiDR1基因的c DNA全长,利用生物信息学分析软件对其基因结构及功能进行分析和预测。再通过qRT-PCR技术对干旱胁迫后的幼苗中的CiDR1表达模式进行研究。[结果]从中间锦鸡儿中克隆到CiDR1基因的c DNA全长共计4 297 bp,Gen Bank登录号为KP277100。生物信息学分析表明,预测的CiDR1蛋白序列中含有1 243个氨基酸残基,具有抗病基因特征结构域TIR、NB-ARC、LRR等,其等电点为6.35,不稳定指数为42.91,不具备信号肽,为非分泌蛋白,定位于细胞质中。定量PCR检测发现,CiDR1基因在幼年期的茎中表达量较低,在成年期的叶片中表达量较高;在干旱胁迫处理后的幼苗中,CiDR1表达水平有明显下降,表明该基因的表达受干旱抑制,可能与中间锦鸡儿适应干旱相关。[结论]中间锦鸡儿在干旱胁迫后其根、茎和叶中CiDR1的表达均明显下降,表明CiDR1的表达受干旱抑制,可能与中间锦鸡儿适应干旱相关,进一步研究发现CiDR1在根、茎、叶中的表达水平可能受发育阶段调控。     

山鸡椒LcGPPS表达模式及其与LcGGPPS蛋白互作分析

[目的]鉴定与山鸡椒精油合成关键酶香叶基二磷酸合酶(LcGPPS)的互作蛋白,为揭示山鸡椒精油主要成分单萜物质的合成机理奠定基础。[方法]通过本地Blast搜索山鸡椒果实转录组数据库,采用RT-PCR克隆山鸡椒GPPS基因(编码异质GPPS小亚基的LcGPPS.SSU1)和山鸡椒GGPPS基因(LcGGPPS1和LcGGPPS3);利用qRTPCR分析其组织特异性表达模式,通过蛋白互作预测和酵母双杂实验验证LcGPPS.SSU1分别与LcGGPPS1和LcGGPPS3的互作关系。[结果]克隆得到LcGPPS.SSU1、LcGGPPS1和LcGGPPS3 3条基因序列,基因表达模式分析表明,LcGPPS.SSU1特异性地在花和果实中高水平表达;蛋白结构互作预测结果显示,LcGPPS.SSU1可以分别与LcGGPPS1和LcGGPPS3互作形成异质二聚体;经过酵母双杂交实验验证发现,仅LcGGPPS3能与LcGPPS.SSU1发生互作。[结论]LcGPPS.SSU1通过与LcGGPPS3蛋白发生互作从而在山鸡椒萜类合成途径中发挥作用,为深入研究山鸡椒萜类合成机制提供参考。     

巨桉EXP基因家族的生物信息学分析

[目的]本研究有助于了解EXP基因家族的基本特征,为深入研究其功能搭建平台。[方法]本研究对从巨桉(Eucalyptus grandis Hill)中筛选出35个EXP基因家族成员(Egr EXP1 Egr EXP35),利用生物信息学方法对其基因特征与表达模式进行综合分析。[结果]巨桉EXP基因分布在8条染色体之上,EXP蛋白均定位在细胞质膜上发挥作用,大多数的家族成员具有信号肽。巨桉EXP编码的蛋白质由α-螺旋、延伸链、无规卷曲、β-转角组成。进化分析结果表明,巨桉EXP蛋白与毛果杨(Populus trichocarpa) EXP蛋白的进化关系接近。35个巨桉EXP基因在巨桉未成熟木质部、成熟叶片、韧皮部、茎尖、木质部以及幼叶组织中表达模式存在显著差异。[结论]EXP基因家族各成员的表达模式不同,Egr EXP17、Egr EXP18可能在巨桉木材形成过程中起重要作用。     

烟草和毛白杨次生维管发育相关MYB转录因子分析

[目的]为探讨额河杨和银灰杨天然杂种的起源机制,[方法]应用18对SSR标记,从分子水平上对新疆额尔齐斯河流域杨属植物的种间关系进行分析研究。[结果]表明:(1)SSR系统发育树将整个流域天然杨属植物分为两大类群,即黑杨派和青杨派为一类,白杨派为一类;(2)白杨派派内系统聚类图显示,银白杨、欧洲山杨、银灰杨三个树种均有较大的遗传分化,特别是杂种银灰杨似乎更大;(3)黑杨派和青杨派的UPGMA分类图显示,青杨派和黑杨派分属于2个分支,其中,青杨派内部分化相对简单,分为2支,均为典型的苦杨;黑杨派内部的分化较为复杂,可分为4类,包括典型的欧洲黑杨、额河杨和回交子代。[结论]杂种额河杨具有更多的欧洲黑杨的遗传成分,因此,将额河杨放到黑杨派是正确的。     

黄河小浪底库区山地栓皮栎人工林土壤呼吸的季节动态

[目的]分离并量化土壤自养呼吸和异养呼吸,探讨各自贡献率及其随季节变化的动态特征。[方法]采用壕沟法和气体红外分析法,研究黄河小浪底库区山地栓皮栎人工林土壤总呼吸、自养呼吸和异养呼吸速率的季节动态变化、贡献率和环境影响因子。[结果]表明:栓皮栎人工林总土壤呼吸、自养呼吸和异养呼吸均呈夏季速率高、冬季速率低。栓皮栎土壤总呼吸、自养呼吸及异养呼吸速率与5 cm土壤温度均呈极显著指数相关,温度敏感性系数Q_(10)值大小为自养呼吸(3.40)异养呼吸(2.90)土壤总呼吸(2.45);栓皮栎土壤总呼吸、自养呼吸、异养呼吸速率与0 10 cm土壤体积含水量均显著线性相关;土壤总呼吸、自养呼吸速率与0 10 cm土壤电导率显著相关。土壤总呼吸和异养呼吸的温度敏感系数Q_(10)值均在冬季最大,夏秋季最小;而自养呼吸的Q_(10)值则呈相反的变化趋势。栓皮栎人工林自养呼吸和异养呼吸对土壤总呼吸的月贡献率为13.23%37.33%和62.67%86.76%,且自养呼吸的贡献率与土壤温度的季节变化规律相似。土壤总呼吸、异养呼吸与自养呼吸的CO2年通量分别为1 616.41、1 199.39、417.02 g·m~(-2)·a~(-1)。[结论]经过区分与定量化土壤总呼吸及其组分,确定异养呼吸为本研究区栓皮栎人工林土壤总呼吸的主要组分,作用于异养呼吸的生物与非生物因子均能显著影响整个森林生态系统表层CO_2总排放通量的大小,进一步为该研究区森林生态系统碳循环与能量流动的进一步量化研究提供参考。     

Relationships between growth and leaf-scale physiological parameters in five Wildstar™ cherry clones (Prunus avium L.)

Infra-red gas exchange analysis was used to measure leaf-scale physiological parameters of five Wildstar™ cherry clones (Prunus avium L.) and two sources of unimproved stock. Assimilation rate (A), evapotranspiration rate (E), stomatal conductance (g _s) and water use efficiency (WUE) were recorded to evaluate whether the quick and simple measurements could be used as a proxy for assessing growth potential of the cherry clones. Differences in A, E and g _s were found between varieties, with clone one always having higher rates than clones two and three. Differences in growth highlighted by an earlier study were linked to the physiological parameters described here. Varieties with high A tended to have good height increment and relative growth rate (RGR), while those with low A, E and g _s had poor height increment. However, clone 5, the clone with the largest height increment, did not have the highest A, suggesting that its good height growth reflected allocation of photoassimilate to main stem growth relative to branches and roots. Likewise, clone 1 and clone 4 (with high A) were ranked only second and third in height growth, indicating that some of the carbon gain was lost in branching. Clones 2 and 3 performed poorly in both physiological parameters and height growth. Although a positive relationship was found between assimilation rate and growth, the relationship was not strong enough to assess growth potential of the cherry clones accurately, perhaps due to differences in the allocation of dry matter within the plant.     

Photosynthetic response to green crown pruning in young plantation-grown Eucalyptus pilularis and E. cloeziana

The loss of foliage through pruning of live branches may reduce tree growth or it may be compensated by photosynthetic up-regulation of the remaining crown. Here, the changes in light-saturated photosynthesis following pruning to remove 50% of green crown length were examined in 4-year-old Eucalyptus pilularis Sm. and Eucalyptus cloeziana F. Muell. trees. The objectives of the study were to: (1) compare leaf-level physiological (light-saturated photosynthesis (A_max), stomatal conductance (g), transpiration (T), dark respiration (R_d), quantum yield (Φ), light compensation point (Γ), water-use efficiency (WUE), nitrogen-use efficiency (NUE)) traits in species with contrasting crown dynamics and structure, (2) examine the effect of crown position on these traits, and (3) examine the effect of pruning on A_max, g, T, WUE, NUE, leaf N and P concentrations and specific leaf area (SLA). Prior to pruning there were no differences in R_d, Γ and Φ between E. pilularis and E. cloeziana but differences in A_max, T, g, leaf N, leaf P, WUE, NUE and SLA. Whereas the rate of physiological processes (A_max, T, and g) and leaf N and P concentrations increased with crown height, R_d, Γ, Φ and SLA declined along this vertical gradient, except in the upper crown of E. cloeziana where A_max, T and g were not different to the lower crown. No up-regulation of photosynthesis or changes in leaf physiology occurred between 6 and 13 months after pruning in either species. The results provide an important basis for modelling pruning effects in process-based tree growth models.     

Toxicity of biocide GCSC-BtA on arthropod pests under different temperature conditions

The present research dealt with the toxicity of GCSC-BtA (Germany–China Scientific Cooperation-Bacillus thuringiensis-Abamectin), a new type biocide developed by conjugating a toxin from Bacillus thuringiensis (B.t.) with Abamectin from Streptomyces avermitilis, on arthropod pests under different temperature conditions. The results showed that GCSC-BtA possessed higher toxicity than B.t. crystal, Abamectin or Cypermethrin, with pest mortalities of 97.9, 93.7, 96.4, 82.3 and 96.7% for Tetranychus cinnabarinus (Boisd.) (Acari, Tetranychidae), Frankliniella occidentalis Pergande (Thys., Thripidae), Aphis fabae Scopoli (Hom., Aphididae), Plutella xylostella (L.) (Lep., Plutellidae) and Cameraria ohridella Deschka et Dimić (Lep., Gracillariidae), respectively. Toxicities of GCSC-BtA to T. cinnabarinus and P. xylostella decreased significantly at the older developmental stage with an exception that egg stage had the highest tolerance, in which LC₅₀s of GCSC-BtA were 0.0001, 0.0019 and 0.0708 mg/ml for nymph, adult and egg of T. cinnabarinus, and that 0.0399, 0.2035 and 0.9033 mg/ml for the 2nd instar larvae, 4th instar larvae and egg of P. xylostella, respectively. Also, the biocide was more effective to the lower stage of A. fabae than the higher one with LC₅₀s of 0.0023, 0.0086 and 0.0171 mg/ml against 1st instar nymph, 3rd instar nymph and adult, respectively. In general, toxicity of the new type of biocide was positively related to temperature against all the tested pests, except against T. cinnabarinus, where it was almost similar at the three temperature conditions. GCSC-BtA displayed significantly lower toxicities at 15/20°C than 20/25°C or 25/30°C against F. occidentalis, A. fabae, P. xylostella and C. ohridella. However, no significant difference was found in the toxicities against F. occidentalis, A. fabae and C. ohridella above 20°C, whereas toxicity against P. xylostella increased as temperature raised. Therefore, to achieve higher efficacy in the field, application rate and time should be devised according to these factors.     

Nitrogen mineralization and maize yields following application of tree prunings to a sandy soil in Zimbabwe

Despite the promotion of prunings as sources of nitrogen for crops, lack of synchronization between N mineralization from prunings and plant uptake remains a major limitation to the impact of prunings on crop yields. A laboratory and a field experiment were therefore carried out to determine the mineralization patterns of selected prunings and assess the potential that exists to improve synchrony by mixing prunings of different quality. The laboratory incubation experiment was conducted for 84 days to determine the C and N release patterns of prunings of different quality and the manipulation of the C and N mineralization trends by mixing prunings of different quality. High quality prunings were considered to be those with high CO₂ evolution rates and mineralize N rapidly. The % C and % N released in 84 days were highest for the high quality prunings of Tithonia diversifolia (70% and 30% respectively) and least for the low quality prunings of Flemingia macrophylla (25% and –5% respectively). The medium quality prunings of Acacia angustissima and Calliandra calothyrsus had similar proportions of released C and N (about 40% C and 10% N). Different mixtures of T. diversifolia with other species showed contrasting influence on C release and N mineralization. Most mixtures released less C than that predicted, but in contrast most mixtures released N at a rate either matching or above the predicted. The % N released was strongly correlated with the polyphenol protein binding capacity (r² = 0.53) and also with the % C released (r² = 0.62). The field experiment was done for two cropping seasons to determine the effects of the prunings of these species and their mixtures on maize grain yield. The prunings were added at 5 t ha^–1 and incorporated into the top 15 cm by hand hoeing in the first season and their residual effects were monitored in the second season. Medium and low quality prunings produced significantly (p < 0.05) higher maize grain yields in the first season ranging between 2.4 t ha^–1 and 3.4 t ha^–1 compared with T. diversifolia which produced 1.7 t ha^–1. This suggested better synchrony in N release and uptake by maize with medium and low quality prunings compared with high quality prunings. The only mixture that indicated improved synchrony was the mixture of T. diversifolia and C. calothyrsus. This study showed that mixing prunings of different quality produce different patterns of N mineralization, some of which were unexpected and had a potential for improving N synchrony.This revised version was published online in November 2005 with corrections to the Cover Date.