Divergent functions of SNAC4–9 and possible mechanisms for tomato adaptation to abiotic stresses

Plants are exposed to all types of abiotic stresses during the process of growth and development, which could adversely affect the productivity and postharvest storage quality of plants. In the current study, the expression of SNAC4–9 and the changes of physiological parameters were analyzed in tomato seedlings under various abiotic conditions and hormone treatments. The results demonstrate that all six genes were induced by these stresses at differential induction levels and that SNAC4–9 gene expression profiles were likely to be related to the ABA, SA, and MeJA signaling pathways. In addition, all of the stress- and hormone-treated seedlings exhibited significant increases in their proline content and antioxidant enzyme activities. MDA was significantly increased in seedlings exposed to stress and decreased in hormone-treated seedlings. These data collectively suggest that SNAC4–9 might function through ABA, SA, and JA signaling pathways and the regulation of proline and antioxidant systems. This study combines molecular biology and physiology to provide valuable information for further exploring the functional roles of NAC genes in response to environmental stresses and indicates that these genes may exhibit potential for enhancing stress tolerance of transgenic tomato plants and further improving the postharvest storage quality of tomato plants.     

DREB1A regulon expression in rd29A:DREB1A transgenic chrysanthemum under low temperature or dehydration stress

Summary

Previously we reported that expression of the Arabidopsis DREB1A gene in chrysanthemum conferred increased tolerance to low-temperature and dehydration stresses, and that transgenic plants in which the DREB1A gene was driven by the abiotic stress-inducible promoter, rd29A, were more tolerant than those plants in which the DREB1A gene was driven by the 35S Cauliflower Mosaic Virus (CaMV) promoter. To understand the molecular basis for this improved tolerance, we isolated 74 DREB1A regulon genes using suppression subtractive hybridisation, then compared their expression patterns in rd29A:DREB1A transgenic plants (rd29A plants) and in 35S:DREB1A transgenic plants (35S plants) under different stress conditions. Our results showed that the increased tolerance to low temperatures and dehydration in rd29A plants was attributed to increased levels of expression of different members of the DREB1A regulon. Levels of expression of 69% or 91% for members of the DREB1A regulon that showed upregulation in rd29A plants were highly correlated with the level of expression of DREB1A in response to low temperature or to dehydration, respectively. These results support the hypothesis that the increased tolerance of rd29A plants to abiotic stresses resulted from elevated expression of the DREB1A regulon.     

Molecular cloning and characterisation of a germin-like protein gene in spinach (SoGLP)

Summary

Germin-like proteins (GLPs) are ubiquitous plant glycoproteins that have been implicated in various plant physiological and developmental processes. In this study, a cDNA clone, designated SoGLP, encoding a germin-like protein from spinach (Spinacia oleracia L.) was isolated and characterised. SoGLP encodes a 208-amino-acid polypeptide with a predicted molecular mass of 22.54 kDa and a pI of 5.95. Phylogenetic analysis indicated that SoGLP belonged to sub-family 3 of the GLP family. Sub-cellular localisation of an SoGLP-GFP fusion protein appeared to detect the protein on the cell walls in transgenic tobacco plants. Real-time quantitative RT-PCR analysis showed that the level of expression of SoGLP in the salt-resistant spinach cultivar (‘Chaoji’) was generally higher than in the salt-sensitive spinach cultivar (‘Daye’) grown in 160 mM nitrate ions for 0.5, 3.0, or 6.0 h. Expression of the SoGLP gene was also induced by other abiotic stresses including polyethylene glycol (PEG), NaCl, salicylic acid (SA), or H₂O₂ treatment. Our results indicate that SoGLP could play important roles during high nitrate stress or under other abiotic stresses.     

枇杷脱水素基因EjDHN5抗低温胁迫功能研究

为研究枇杷(Eriobotrya japonica Lindl.)脱水素基因(Ej DHN5)在枇杷抗低温胁迫中的功能,将其在烟草中进行过量表达,其中表达水平比较高的两个株系L24和L26被用来进行功能研究。野生型和转基因烟草苗分别用0、2、4和8μmol·L~(-1)的甲基紫精(Methyl viologen,自由基发生剂)处理并进行光照培养,结果显示,4和8μmol·L~(-1)甲基紫精处理使野生型烟草苗存活率分别下降到73.6%和56.9%,光系统Ⅱ(PSⅡ)活性显著降低,而转基因苗的存活率仍维持在100%,PSⅡ活性明显高于野生型。对六叶期烟草的叶盘进行甲基紫精处理,野生型烟草叶盘的叶绿素含量显著低于转基因株系,而活性氧含量和MDA含量明显高于转基因株系。对野生型和转基因烟草苗进行低温处理,转基因株系生长状况明显优于野生型,活性氧含量低于野生型,膜质过氧化程度轻于野生型。这些结果表明,过量表达EjDHN5能提高烟草的抗氧化胁迫和抗低温胁迫能力,推测EjDHN5在提高枇杷抗低温能力方面起着重要作用可能与其能提高抗氧化胁迫能力有关。     

Effects of proline on antioxidant system in leaves of grapevine (Vitis vinifera L.) exposed to oxidative stress by H2O2

Although proline is one of the major computable organic solutes that accumulate in many plant species in abiotic stresses, a hot debate continues about whether proline accumulation is a reaction to abiotic stress, or a plant's response is associated with stress tolerance. The effects of proline on antioxidative system in grape leaves of Vitis vinifera L. cv., ‘Öküzgözü’ exposed to oxidative stress by H₂O₂ was investigated. Endogenous proline, hydrogen peroxide (H₂O₂), malondialdehyde (MDA) concentrations, percentage of electrolyte leakage (EL), and some of the antioxidant enzyme activities; such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and guaiacol peroxidase (POD) were measured spectrophotometrically. Inhibitory effect of H₂O₂ on antioxidant enzyme activities, MDA, and EL was found. In the presence of proline, SOD and CAT activities decreased, while POD and APX activities increased. Proline pre-treatment resulted in a decrease in cellular H₂O₂ content, MDA, and EL, while cellular concentration of proline increased. Based on the finding, it was suggested that proline and H₂O₂ could play an important role in oxidative stress injury of grapevine leaves grown in vitro culture. Also, proline might have a direct positive effect on antioxidant enzyme system, membrane phase change, MDA, and EL.     

转香樟CcCBFs烟草的抗逆性分析

为探究香樟CBF基因（CcCBFa、CcCBFb和CcCBFc）增强植株非生物胁迫抗性的功能,对获得的T1代转基因烟草,分别进行300 mmol · L-1甘露醇模拟干旱、300 mmol · L-1 NaCl溶液模拟高盐和4 ℃模拟低温胁迫处理,测定其丙二醛、脯氨酸和可溶性糖含量,探讨其抗逆性。结果表明：CcCBFb和CcCBFc可显著增强烟草抗旱性;CcCBFa和CcCBFc可显著增强烟草抗盐性;CcCBFc可显著增强烟草抗寒性。CcCBFc可以提高转基因烟草抗旱、抗盐、抗寒能力。     

Current trends in the genetic engineering of vegetable crops

Vegetables play an important role in human nutrition and health. Cultivation of vegetable crops is an integral part of the agricultural economy of many developing countries. Vegetable crop productivity and quality are seriously affected by several biotic and abiotic stresses, which destabilize rural economies in many countries. Moreover, absence of proper post-harvest storage and processing facilities leads to qualitative and quantitative losses. In the past four decades, conventional breeding has contributed significantly for the improvement of vegetable yields, quality, post-harvest life, and resistance to biotic and abiotic stresses. However, there are many constraints in conventional breeding, which can only be overcome by advancements made in modern biology. In the last decade various traits such as biotic stress resistance, quality and storage life have been successfully engineered into vegetable crops and some of them have been commercialized. In recent years significant progress has been made to manipulate vegetable crops for abiotic stress tolerance, quality improvement and pharmaceutical and industrial applications. Although the progress in commercialization of transgenic vegetable crops has been relatively slow, transgenic vegetables engineered for nutraceutical and pharmaceutical use will contribute significantly to the value added agriculture in near future.     

Induction of drought tolerance by salicylic acid in seedlings of cucumber (Cucumis sativus L.)

Summary

Salicylic acid (SA) is a common, plant-produced signal molecule that is responsible for inducing tolerance to a number of biotic and abiotic stresses. An experiment was therefore conducted to test whether the application of SA at various concentrations (0, 0.25, 0.50, 0.75, or 1.00 mM) by seed soaking, or as a foliar spray would protect cucumber (Cucumis sativus L.) seedlings subjected to drought stress. Thirty-six-day-old seedlings (n = 12 seedlings per treatment) were exposed to drought stress for 14 d. Pretreatment with SA improved the majority of the physiological (e.g., relative chlorophyll content and chlorophyll fluorescence ratio) and morphological parameters (e.g., shoot and root fresh and dry weights) measured in cucumber seedlings subjected to drought stress. SA ameliorated the injuries caused by drought stress by increasing shoot tissue proline contents and preventing an increase in leaf electrolyte leakage. SA was more effective at increasing the drought tolerance of cucumber seedlings when applied using the seed-soaking method, rather than as a foliar spray. The best drought protection appeared to be obtained when seeds were soaked in 0.50 mM SA.     

Isolation and expression analysis of differentially expressed genes in stem tissue of the Greek lemon cv. Adamopoulou

Lemon (Citrus limon (L.) Burm. f.) is susceptible to mal secco, a serious vascular disease caused by the fungus Phoma tracheiphila (Petri) Kant. and Gik., as well as low temperatures. The greek lemon cultivar Adamopoulou, thought to be derived from the Portuguese cultivar Lisbon, exhibits enhanced resistance to mal secco and cold as opposed to cv. Lisbon. Suppression subtractive hybridization (SSH) was employed for the isolation of differentially expressed genes in lemon stem tissue. A subtractive cDNA library was constructed and a total of 296 clones were sequenced. The obtained sequences were edited, resulting in 56 non-redundant ESTs. Sequence analysis revealed homology to previously identified genes involved in defense mechanisms against biotic and abiotic stresses, as well as sequences with no significant similarity in the GenBank. Selected ESTs were analyzed by real-time PCR for confirmation purposes. This analysis revealed significant expression differences between the two cultivars for genes expressing allantoinase, ultraviolet-B-repressible protein, 4-coumarate:CoA ligase and other proteins that are known to be upregulated under biotic and abiotic stress conditions.     

过表达香樟CcCBFb的烟草非生物胁迫抗性增强

为探究香樟Cc CBFb基因增强植株非生物胁迫抗性的功能,通过农杆菌介导法将该基因转入烟草中,经PCR和半定量RT-PCR技术鉴定阳性转基因株系后,对获得的T1代转基因无性系(T2和T4株系)以及野生型(WT)进行干旱(0、150、300和450 mmol·L~(-1)的甘露醇)、高盐(0、100、200和300 mmol·L~(-1)的Na Cl)、4℃低温、–4℃冰冻胁迫处理,结果显示:转基因烟草在干旱和高盐胁迫下,幼苗的存活率均高于野生型;经4℃低温处理后,转基因烟草植株的脯氨酸和可溶性糖含量均显著高于野生型植株,而丙二醛(MDA)含量均低于野生型植株;经–4℃的冰冻处理6 h后,野生型和转基因T4株系植株叶片均出现不同程度的萎蔫,而转基因T2株系植株未出现不良现象。由此可见,过表达CcCBFb基因不但能够增强烟草的抗旱和抗盐性,而且能够显著增强抗寒性。     

Paclobutrazol application enhances antioxidant enzyme activities in pomegranate plants affected by cold stress

Paclobutrazol (PBZ) is a member of the triazole plant growth inhibitor group that is responsible for inducing tolerance to a number of biotic and abiotic stresses. An experiment was, therefore, conducted to test whether PBZ application at various concentrations (0, 25, 50, or 75 mg L^?1) through foliar spray or soil drench would protect pomegranate (Punica granatum) seedlings cv. Robab, subjected to freezing at ?3°C for 7 hours. PBZ improved the growth rate of pomegranate seedlings subjected to freezing stress and increased relative leaf chlorophyll content, chlorophyll fluorescence ratio, relative water content, soluble carbohydrate content, and enzyme activity of ascorbate peroxidase and guaiacol peroxidase compared with the control at the end of experiment. PBZ ameliorated the injury caused by freezing stress by inhibiting increases in proline content and leaf electrolyte leakage, which suggested that PBZ ameliorated the negative effect of freezing stress. The best protection was obtained from plants treated with PBZ at 50 or 75 mg L^?1. The most evaluated parameters were not affected by PBZ application method.     

转沙冬青锌指蛋白基因AmZFPG 烟草非生物胁迫抗性分析

 分析了沙冬青（Ammopiptanthus mongolicus）锌指蛋白基因AmZFPG 在非生物胁迫下的表达 特性,结果显示AmZFPG 受低温、干旱、高盐胁迫诱导表达,表明该蛋白参与多种胁迫相关的信号转导 和应答反应。为进一步探索AmZFPG 的功能,构建了真核表达载体AmZFPG-pCAMBIA2300,将该基因 转入烟草（Nicotiana tabacum L.）,对转基因烟草T1 代进行非生物胁迫分析,结果显示,转基因烟草的耐 寒性、耐旱性、耐盐性均获得了提高。     

苹果MdCBL家族基因表达分析及MdCBL1的功能鉴定

利用生物信息学的方法,对10个苹果MdCBLs家族蛋白的功能域进行预测,并与拟南芥AtCBLs家族的10个蛋白进行了系统进化分析,同时对苹果MdCBLs基因进行了系统的命名。利用qRT-PCR,检测了苹果MdCBLs基因在不同组织的表达及对非生物胁迫(包括盐、低温、ABA、干旱)的响应,结果表明,MdCBLs在苹果不同组织及非生物胁迫中起重要作用。另外,通过遗传转化苹果愈伤组织鉴定了MdCBL1在盐胁迫中的功能,MdCBL1过量表达明显提高了转基因愈伤组织的盐胁迫抗性。