Isolation of high quality and polysaccharide-free DNA from leaves of Dimorphandra mollis (Leguminosae), a tree from the Brazilian Cerrado

Dimorphandra mollis (Leguminosae), known as faveiro and fava d'anta, is a tree that is widely distributed throughout the Brazilian Cerrado (a savanna-like biome). This species is economically valuable and has been extensively exploited because its fruits contain the flavonoid rutin, which is used to produce medications for human circulatory diseases. Knowledge about its genetic diversity is needed to guide decisions about the conservation and rational use of this species in order to maintain its diversity. DNA extraction is an essential step for obtaining good results in a molecular analysis. However, DNA isolation from plants is usually compromised by excessive contamination by secondary metabolites. DNA extraction of D. mollis, mainly from mature leaves, results in a highly viscous mass that is difficult to handle and use in techniques that require pure DNA. We tested four protocols for plant DNA extraction that can be used to minimize problems such as contamination by polysaccharides, which is more pronounced in material from mature leaves. The protocol that produced the best DNA quality initially utilizes a sorbitol buffer to remove mucilaginous polysaccharides. The macerated leaf material is washed with this buffer until there is no visible mucilage in the sample. This protocol is adequate for DNA extraction both from young and mature leaves, and could be useful not only for D. mollis but also for other species that have high levels of polysaccharide contamination during the extraction process.     

中国沙棘HrANR基因及黄酮类累积与抗旱的关系

花青素还原酶(anthocyanidin reductase, ANR)是合成黄酮类物质的关键酶之一,为明确其编码基因结构及干旱胁迫下的表达模式和黄酮类物质含量及二者之间的相关性,该文从中国沙棘转录组数据中筛选获得1个ANR基因,命名为HrANR基因。采用生物信息学软件对基因序列及编码蛋白进行分析,并对不同胁迫下各组织中HrANR基因的表达量和叶中黄酮类化合物含量进行相关性分析。结果表明:(1)中国沙棘HrANR基因ORF为1 017 bp,编码338个氨基酸,为稳定的亲水性蛋白,其ANR同源蛋白具有明显的科属特性。(2)干旱胁迫下HrANR基因在中国沙棘根、茎、叶中均有表达,但表达趋势不同,其中在根中的表达呈先升高后降低再升高的趋势,在茎中呈持续下降的趋势,在叶中呈先升高后持续降低的趋势。(3)通过芦丁标准曲线获得不同胁迫程度下中国沙棘叶内黄酮类的含量,表明黄酮类含量呈先持续上升,随后略有下降,复水后上升至最高点的变化趋势,表明干旱胁迫初期叶黄酮类含量与干旱胁迫呈正相关,在严重胁迫下黄酮类含量与胁迫呈负相关。(4)叶和茎的HrANR基因表达量与黄酮类含量呈负相关(P_叶=-0.751,P_茎=-0.934),根中呈正相关(P_根=0.444)。综上表明,中国沙棘HrANR基因的表达及黄酮类含量变化与其抗旱性密切相关,其结果为中国沙棘抗旱机制的阐明提供了依据。     

Impacts of simulated drought stress and artificial damage on concentrations of flavonoids in <Emphasis Type="Italic">Jatropha curcas</Emphasis> (L.), a biofuel shrub

We studied the possible roles of flavonoids in the antioxidant and antiherbivore chemistry in Jatropha curcas (L.), a Latin American shrub that holds great potential as a source of biofuel. Changes in flavonoid concentrations in the leaves of J. curcas seedlings exposed to artificial damage and to different rainfall patterns were assessed by applying a 3²-factorial experiment in a greenhouse. The concentrations of different flavonoids in the leaves of seedlings were significantly affected by interaction effects of artificial damage, drought stress and age of the seedling. The highest flavonoid concentrations were obtained in seedlings imposed to the highest percentage of artificial damage (50 %) and grown under extreme drought stress (200 mm year^?1). In this treatment combination, flavonoid concentrations were three-fold as compared to seedlings exposed to the same level of artificial damage but grown in 1900 mm year^?1 rainfall application. Without artificial damage, the concentration of flavonoids in the seedlings grown in 200 mm year^?1 rainfall application was still two-fold compared to seedlings grown in higher (>800 mm year^?1) rainfall applications. Thus, the observed flavonoid concentration patterns in the leaves of J. curcas seedlings were primarily triggered by drought stress and light rather than by artificial damage, suggesting that drought causes oxidative stress in J. curcas.     

干旱胁迫下红砂幼苗非结构性碳水化合物动态变化特征

该试验以荒漠区主要建群种红砂幼苗为研究对象,设置适宜水分(CK)、轻度干旱(MD)、中度干旱(SD)和重度干旱(VSD)4个胁迫处理(即田间持水量的80%、60%、40%和20%),采用盆栽控水试验,分别测定干旱胁迫15、30、45和60 d时红砂幼苗的叶、茎、粗根和细根中非结构碳水化合物(NSC)及其组分的含量,分析不同胁迫强度下不同干旱持续时间红砂幼苗NSC的动态变化及各组分差异,以揭示红砂NSC对干旱胁迫的响应机制。结果表明:(1)干旱胁迫强度和胁迫持续时间对红砂幼苗不同器官NSC及其组分均有显著影响,其中胁迫持续时间对NSC动态变化的影响尤为显著。(2)干旱胁迫初期,红砂叶中的NSC含量呈下降趋势,而茎中的NSC含量呈上升趋势,粗根和细根中NSC含量在各胁迫处理下基本保持稳定。(3)干旱胁迫后期,红砂叶和茎中的可溶性糖、淀粉和NSC含量逐渐增加,而粗根和细根中的淀粉和NSC含量呈下降趋势(中度干旱除外),且这一时期重度干旱处理下各器官可溶性糖和NSC的含量明显高于CK。研究发现,重度干旱胁迫能显著诱导提高红砂幼苗不同器官中的NSC含量,并通过分解根中淀粉和增加叶片中可溶性糖含量的方式来调节细胞渗透势平衡,以维持细胞活力,进而保持红砂在干旱胁迫后期的存活。     

Changes in the composition of phenolic compounds and antioxidant properties of grapevine roots and leaves (Vitis vinifera L.) under continuous of long-term drought stress

Grapevine seedlings Vitis vinifera L. were grown in a greenhouse under optimum conditions (soil moisture ca 70 %) and under drought stress (soil moisture ca 30 %). Drought stress caused reduction in total phenolic compounds in grapevine leaves and roots, where were identified tree phenolic acids: caffeic acid, p-coumaric acid and ferulic acid. All acids found in leaves and roots occurred in the ester-bound form. Only caffeic acid in leaves appeared in the free and ester-bound form. Caffeic acid was present in the highest concentrations. The content of ferulic acid was the lowest in both tissues. The levels of all phenolic acids in leaves and roots decreased significantly under the drought stress. All the extracts from grapevine leaves and roots had antioxidative properties, but the antiradical activity of the extracts obtained from roots subjected to drought stress was lower to the control. The results of the analysis revealed that long-term drought stress caused a decrease in selected elements of secondary metabolism in such a different plant tissues that are the leaves and roots of the grapevine.     

盐分和干旱对沙枣幼苗生理特性的影响

以沙枣(Elaeagnus angustifolia L.)幼苗为实验材料,分别对其进行轻度干旱(土壤含水量7%—9%)、重度干旱(土壤含水量3%—5%)、100 mmol/L NaCl以及100 mmol/L NaCl处理下不同程度的盐旱共胁迫处理,处理2周后测其生理指标,包括生长指标、光合指标、渗透调节指标以及复水后生长指标,研究盐旱共胁迫对沙枣幼苗生理特性的影响。结果表明:和对照相比,轻度干旱对沙枣幼苗的生物量没有显著影响,重度干旱处理明显降低了沙枣的生物量,无论是轻度干旱还是重度干旱,都显著降低了沙枣幼苗的净光合速率、K~+含量,显著增加了Na~+含量、脯氨酸含量、可溶性糖含量、有机酸含量、总酚和类黄酮含量;和对照相比,100 mmol/L NaCl处理显著降低了沙枣幼苗的生物量、净光合速率和K~+含量,显著增加了Na~+含量、脯氨酸含量、可溶性糖含量、有机酸含量、总酚和类黄酮含量;和盐处理相比,轻度干旱和盐分共胁迫对沙枣幼苗的各项指标没有显著差异,而重度干旱和盐分共胁迫明显降低了沙枣幼苗的生物量、净光合速率;复水一周后,只有轻度干旱可以回复到对照水平。以上结果表明,盐分和干旱处理明显抑制了沙枣幼苗的生长,轻度干旱和盐分共胁迫条件下,沙枣幼苗表现出一定的交叉适应现象,而重度干旱却加重了盐害。     

两种沙生植物抗旱生理指标的比较研究

以沙坡头地区自然生长和人工灌溉条件下的沙冬青（Ａｍｍｏｐｉｐｔａｎｔｈｕｓ ｍｏｎｇｏｌｉｃｕｓ）和柠条（Ｃａｒａｇａｎａ ｋｏｒｓｈｉｎｓｋｉｉ）为材料,对它们部分抗旱生理指标的变化进行了研究,结果表明,沙冬青对干办迫最敏感,这些生理指标随着个体发育阶段的不同变化,并具种间差异性。可溶性糖和游离氨基酸（特别是Ｐｒｏ）在干旱环境植物中均有较高含量,沙冬青有较高的热值,类黄酮水平未见差异性变化。     

干旱胁迫对科尔沁沙地榆树幼苗C、N、P化学计量特征的影响

设置适宜水分、轻度、中度和重度干旱胁迫处理(即80%、65%、50%和35%田间持水量),研究干旱胁迫对榆树幼苗不同器官C、N、P化学计量的影响.结果表明:与适宜水分处理相比,干旱胁迫导致C含量在叶、茎、粗根和细根中增加,N含量在叶、茎和粗根中增加,P含量在叶、茎和粗根中下降,在细根中增加,C:N在叶和茎中下降,C:P及N:P在叶、茎和粗根中增加,在细根中下降.各器官间C含量呈显著正相关,而各器官间N与P相关性均不显著.土壤含水量与各器官C含量,叶N含量,细根P含量及C:N,叶、茎和粗根C:P及N:P呈显著负相关,而与细根N含量,叶、茎和粗根P含量,细根C:P及N:P呈显著正相关.因此,干旱胁迫影响榆树幼苗对N、P的吸收及向上运输,幼苗生长主要受N限制;随着干旱胁迫的加剧,叶、茎和粗根生长受P限制作用增强.     

干旱胁迫对烟草两品种幼苗生理生化特性及NtDEGP5基因表达的影响

以烟草Nicotianatabacum抗旱品种NC82和干旱敏感品种云烟87幼苗为材料,利用PEG-6000对幼苗进行干旱胁迫处理,研究干旱胁迫对烟草不同品种Fv/Fm、SOD活性、POD活性、MDA含量等生理生化指标及NtDEGP5基因表达的影响,为进一步开展NtDEGP5基因的功能研究提供依据。结果表明,干旱胁迫对烟苗的Fv/Fm、SOD活性、POD活性、MDA含量有影响,随着干旱的持续,Fv/Fm呈逐渐下降趋势,POD、SOD活性和MDA含量呈先升高后降低的趋势;不同品种对干旱胁迫的响应有差异,在干旱胁迫0～9 h,云烟87幼苗的Fv/Fm降幅大于NC82,POD、SOD活性、MDA含量的增幅小于NC82,胁迫12h后与对照差异不显著。干旱胁迫下不同品种、不同器官间的Nt DEGP5基因表达有差异,云烟87幼苗根、茎、叶中的表达量均较低,NC82幼苗根中的表达量较低,但干旱胁迫9～12 h后叶和茎的表达量较高。NC82幼苗叶片NtDEGP5基因表达量与Fv/Fm、POD活性、MDA含量呈显著正相关。Fv/Fm、SOD活性、POD活性、MDA含量可作为烟草苗期抗旱性评价指标,NtD...     

Changes in Eucalyptus camaldulensis essential oil composition as response to drought preconditioning

Abstract

Water deficit, a common constraint in forestry, is the main cause of plant stress during plantation. The survival and growth of seedlings is also compromised by herbivory during establishment. The potential of nursery preconditioning to enhance survival chances of future trees by reducing palatability or attracting beneficial insects as a result of changes in chemical defences may be an answer to overcome this situation. Changes in essential oil production and composition were evaluated by GC and GC-MS in Eucalytus camaldulensis seedlings submitted to drought during four weeks at the last stage of nursery period (20 weeks). Significant changes in essential oil relative composition were found. Seedlings (young leaves) submitted to drought developed a terpenoid blend, which has been previously associated to mature leaves and related to their higher resistance towards herbivory. Total amount of non-oxygenated terpenes decreased by 44%, whereas some oxygenated sesquiterpenes (globulol, epiglobulol and ledol) were doubled, and 1,8-cineole content was enhanced by 28.3%.     

土壤自然干旱处理对钩藤生长与生理特征及主要药用成分积累的影响

以一年生钩藤实生苗为试材,通过连续土壤控水12 d盆栽试验,研究持续性土壤自然干旱对钩藤幼苗生长、抗逆生理指标及其主要药用成分含量的影响。结果表明:(1)随着干旱胁迫时间的延长,钩藤根和茎叶生物量以及叶片相对含水量(RWC)显著持续下降(P0.05),而根冠比、叶片丙二醛(MDA)含量及相对电导率(REC)逐渐升高。(2)随着干旱胁迫时间的延长,钩藤叶片叶绿素a、b含量先增高后下降,叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性先上升后下降,且POD活性最先达到峰值,CAT活性增幅最大;叶片中脯氨酸(Pro)、可溶性糖(SS)和可溶性蛋白(SP)含量逐渐升高,且Pro表现出更强的渗透调节能力。(3)钩藤幼苗叶片、主茎和带钩茎枝中钩藤碱与异钩藤碱含量随着干旱胁迫时间的延长而呈先增高后下降的趋势,响应时间先后顺序依次为叶片、主茎、带钩茎枝,生物碱含量由高到低依次为带钩茎枝、叶片、主茎。研究发现,在土壤持续自然干旱条件下,钩藤幼苗生长受到一定影响,但植株能通过提高其抗氧化酶活性和积累渗透调节物质来提高吸水和保水能力,有效抵御干旱逆境;且土壤自然干旱胁迫4～8 d有利于主要药效成分钩藤碱与异钩藤碱的积累;土壤相对含水量在42%～53%时,钩藤幼苗耐旱性较强且钩藤碱与异钩藤碱含量较高。     

等渗的盐分和水分胁迫对芦荟幼苗生长和离子分布的效应

 研究了等渗透势（-0.44、-0.88 MPa）NaCl和PEG 6000处理对六叶龄芦荟(Aloe vera)幼苗叶片生长速率、干物质积累、电解质渗漏和离子吸收、分配的效应。结果表明: -0.44、-0.88 MPa NaCl和PEG处理10 d均明显抑制芦荟幼苗叶片伸长生长,植株干物质积累速率显著降低, 叶片含水量降低,叶片细胞电解质渗漏率上升。NaCl对芦荟幼苗生长的抑制作用显著大于PEG处理的。不同器官离子含量、根系和叶片横切面X-射线微区分析结果表明, NaCl胁迫导致芦荟体内Na+、Cl－含量显著上升,根中增幅明显高于叶片,其中Cl－尤为显著。NaCl胁迫严重抑制芦荟对K+ 和Ca2+ 的吸收及其向叶片的运输,根、叶K+/Na+、Ca2+/Na+ 比率显著下降,而PEG胁迫对离子平衡的干扰较轻,是芦荟对水分胁迫的适应能力高于盐胁迫的主要原因之一。但芦荟对 -0.44～-0.88 MPa NaCl胁迫仍有一定的适应能力,主要原因是：1) 根系对离子的选择性吸收和运输较强,并随着盐胁迫强度增加其选择性增强; 2) 芦荟叶片中的盐分在贮水组织中显著积累,明显高于其它组织细胞。同时,芦荟是CAM（景天酸代谢）途径植物,蒸腾极小,盐分随蒸腾流进入地上部的机会小。     

CCC提高花生幼苗抗旱性的研究

用75、150、300ppm CCC 处理三叶期花生(Arachis hypogaea L.)幼苗,可减低干旱时期花生幼苗叶片质膜透性和叶片含水量的降低程度。150ppm CCC 能提高幼苗过氧化物酶活性,减小丙二醛的生成速率和超氧歧化酶的破坏程度。CCC 处理的花生幼苗,可提高在正常生长条件和干旱时期幼苗叶片内源脱落酸(ABA)水平,以150 ppm 效果最好。CCC 提高花生幼苗抗旱性的原因可能在于诱导幼苗内源 ABA 水平的提高,从而影响幼苗体内有关代谢反应。     

干旱胁迫条件下AMF促进小马鞍羊蹄甲幼苗生长的机理研究

采用温室水分控制试验,在干旱胁迫条件下,定量化研究优势丛枝菌根真菌(AMF)影响优势乡土植物小马鞍羊蹄甲(Bauhinia faberi var.microphylla)幼苗生长的机理,主要通过研究干旱胁迫条件下摩西球囊霉菌(Funneliformis mosseae)与小马鞍羊蹄甲的共生关系,阐明AMF在植物生长初期的作用。结果表明,干旱胁迫条件下,摩西球囊霉菌能够很好地侵染幼苗,侵染率高达89%—97%,并且不受水分条件影响。接种的幼苗最大光合速率、水分利用效率随着干旱胁迫程度从重度到轻度(水分从低到高)逐渐增大,相反地,叶片脯氨酸含量逐渐减小。接种显著地促进幼苗株高、叶片数、叶面积、根长、根面积等生长指标,提高幼苗各部分生物量、地上地下磷(P)含量。当含水量为60%田间持水量时,AMF促进小马鞍羊蹄甲幼苗吸收P的效果最好。接种还显著影响幼苗的生物量分配,在重度干旱胁迫时影响P分配,水分条件也显著影响幼苗的生物量分配。此外,接种和水分的交互作用对叶生物量、总生物量、生长指标以及地上部氮(N)总量影响显著。结果表明干旱胁迫条件下菌根效应显著,并在干旱条件下显著促进了小马鞍羊蹄甲幼苗的生长,这为进一步干旱河谷植被恢复提供了理论依据。     

岳桦幼苗光合特性和非结构性碳水化合物积累对干旱胁迫的响应

以长白山林线树种岳桦为对象,利用生长控制试验进行干旱处理,研究干旱对岳桦幼苗光合特性及非结构性碳水化合物(NSC)积累的影响.结果表明:干旱显著降低了岳桦幼苗的净光合速率和气孔导度,提高了其水分利用效率;干旱显著增加了岳桦幼苗叶、皮、干和根中的可溶性糖和总NSC的含量,但显著降低了淀粉含量;随着干旱的持续,叶片的气孔导...     

干旱胁迫下小麦幼苗根、叶多胺含量和多胺氧化酶活性的变化

 以抗旱性不同的两个小麦品种（‘晋麦33’和‘温麦8’）（Triticum aestivum cv. Jinmai 33 and Wenmai 8）为材料,研究了干旱胁迫下多胺含量和多胺氧化酶活性的变化。结果表明：旱过程中,幼苗根、叶中腐胺（Put）、亚精胺（Spd）、精胺（Spm）3种多胺含量和多胺氧化酶（PAO）活性先迅速升高,而后下降。与抗旱性弱的‘晋麦33’相比,抗旱性强的品种‘温麦8’幼苗根、叶中Spd、Spm 含量初期升高幅度大,之后下降速率减慢;PAO活性的变化与之相反,‘晋麦33’的PAO活性提高的幅度大于‘温麦8号’。多胺含量和PAO活性在小麦幼苗的根与叶之间呈极显著正相关。干旱初期,小麦根、叶中多胺迅速积累可能是干旱胁迫反应的一个信号,随后较高的Spd、Spm 水平有利于增强小麦幼苗的抗旱性。     

旱生和湿生生境对蒲公英体内抗氧化物质的影响

选择旱生和湿生生境中生长的蒲公英(Taraxacum mongolicum Hand.-Mazz)分根、叶、花序分别测定抗氧化成分及总抗氧化能力,并比较了SOD同工酶谱的变化情况。结果表明不同生境选取的蒲公英叶片中抗氧化物质以SOD、POD、CAT等抗氧化酶和Vc等小分子为主,花序中可溶性糖、类黄酮和绿原酸含量最高;湿生蒲公英各器官尤其是根中的SOD、CAT、POD活性高于旱生蒲公英对应器官中的酶活性,旱生蒲公英各器官尤其是花序中的可溶性糖、类黄酮和绿原酸含量高于湿生蒲公英对应器官中的含量;旱生和湿生生境未诱导出新的SOD同工酶带;旱生和湿生蒲公英对应器官的总抗氧化能力接近。     

干旱胁迫下三种三角梅的生理反应

该研究对勤花三角梅(Bougainvillea buttiana‘Miss Manila’)、樱花三角梅(B.glabra‘Imperial Delight’)和金心鸳鸯三角梅(B.peruviana‘Thimma’)三个三角梅品种的幼苗进行人工干旱胁迫,分析了干旱胁迫对三角梅品种的外观形态和生理生化水平的影响,探讨了不同品种三角梅对干旱胁迫的适应能力。结果表明:随着干旱时间的延长,三角梅品种的叶片出现下垂、皱缩、变黄、脱落,最后植株死亡;过氧化物酶POD活性先升高后降低,而超氧化物歧化酶SOD活性则逐渐降低,降低速度因品种而异;qP值降低、qN值升高、PSⅡ反应中心光化学效率(Fv/Fm)、PSⅡ潜在活性(Fv/Fo)均呈现降低。该研究结果为三角梅的栽培应用和品种选择提供了依据。