Class II chitinase accumulated in the bark tissue involves with the cold hardiness of shoot stems in highbush blueberry (Vaccinium corymbosum L.)

To understand the development of cold hardiness in highbush blueberry plants (Vaccinium corymbosum L.), we investigated seasonal changes in the protein compositions of current-year and overwintered shoots. Electrophoresis using SDS-polyacrylamide gels indicated that the amounts of a few proteins increased in autumn, in association with an enhancement of cold hardiness. Of these proteins, 65- and 60-kDa proteins were confirmed to be dehydrins by western blotting. While the levels of most of the accumulated proteins decreased in April, a 27-kDa protein maintained its level in the overwintered stem during spring. The amino acid sequence deduced from a cDNA for this protein showed significant similarities with known chitinases. The stem segments from overwintered shoots sampled in July showed higher cold hardiness against sub-zero temperature than those from current-year shoots, and the bark tissue of overwintered shoots had higher cold resistance than that of current-year shoots. The 27-kDa protein (BC27) was mostly found in the bark tissue of stems. Appreciable accumulation of this protein in floral buds was not detected. The purified BC27 protein exerted a cryoprotective effect on protoplasts subjected to a freeze–thaw treatment. These results suggest that the stems of highbush blueberry develop a system of cold hardiness different from that of the buds and apices, and that the accumulation of chitinase is involved in tolerance to low temperature in winter or unseasonably low temperature in spring.     

Molecular cloning and functional analysis of a flavanone 3-hydroxylase gene from blueberry

Vaccinium corymbosum (blueberry) is touted as a superfood with numerous health benefits due to its high levels of flavonoids. Flavanone 3-hydroxylase (F3H) is a key regulatory enzyme of the flavonoid pathway. In this study, we cloned the full-length cDNA of F3H (designated VcF3H) from young blueberry leaves using rapid amplification of cDNA ends (RACE). The cDNA contained a 1080-bp open reading frame that encoded a 359-amino acid protein. The deduced VcF3H protein showed high similarities to other plant F3Hs. Conserved amino acid motifs required for ferrous iron binding (HXD) and 2-oxoglutarate binding (RXS) were identified in VcF3H, VcFLS (flavonol synthase), and VcANS (anthocyanidin synthase). Quantitative RT-PCR analysis demonstrated that VcF3H was expressed in all tissues tested, with particularly high expression in young leaves, fruits (pink and blue), and stems. Anthocyanins accumulated mainly in fruits, whereas flavonols were found mainly in leaves and stems. Furthermore, the expression pattern of VcF3H was similar to that of VcCHS, VcDFR, and VcANS in various tissues. Heterologous expression of VcF3H in Arabidopsis thaliana increased the anthocyanin content in leaves, but did not affect the flavonol content. Thus, VcF3H seems to be involved in anthocyanin synthesis in the flavonoid biosynthetic pathway when ectopically expressed in Arabidopsis.     

Changes in nutrient concentrations and leaf gas exchange parameters in banana plantlets under gradual soil moisture depletion

Changes in mineral nutrient concentration, growth, water status and gas exchange parameters were investigated in young banana plants (Musa acuminata cv. ‘Grand Nain’) subjected to gradual soil moisture diminution. Experiments were performed in glasshouse under controlled temperature, and water stress was imposed by ceasing irrigation for 62 days. The data showed a parallel decrease of leaf gas exchange parameters and soil moisture initiated few days after the imposition of water stress. However, the leaf relative water content (RWC) showed a minor decrease in response to drought. The onset of growth reduction evaluated as plant height, pseudostem circumference, number of newly emerged leaves, leaf area, and leaf and root biomass took place approximately between 34 and 40 days after the beginning of the stress period. In addition, drought did not modify nitrogen and phosphorus concentrations in foliar and root tissues; however, it increased potassium, calcium, magnesium, sodium and chloride in leaves, and only calcium, sodium and chloride in roots. Collectively, the data reveal that banana plants show a drought avoidance mechanism in response to water stress. After a prolonged drought period, leaf RWC was hardly reduced, while gas exchange and growth parameters were reduced drastically. Increasing leaf mineral concentration could have help to maintain leaf RWC due to osmotic adjustment mechanism.     

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。     

Photosynthetic characteristics of highbush blueberry (Vaccinium corymbosum cv. Bluecrop) leaves in response to water stress and subsequent re-irrigation

SUMMARY

Gas exchange and photosystem II (PSII) activities in the leaves of 2-year-old ‘Bluecrop’ highbush blueberry (Vaccinium corymbosum) were monitored during water stress and subsequent re-irrigation to investigate the effects of the intensity of water stress on changes in photosynthetic characteristics. The blueberry shrubs were not irrigated for 3 to 5 weeks, then re-irrigated daily up to 8 weeks. The decrease in soil water potential during water stress caused a progressive decrease in leaf water potential. Soil water potentials decreased to -0.26 MPa and -0.34 MPa at 3 and 5 weeks, respectively, following water stress, but recovered following subsequent re-irrigation, while the soil water potential in daily-irrigated shrubs was maintained at over -0.13 MPa throughout the experiment. Chlorophyll concentrations decreased with an increasing duration of water stress. Chlorophyll concentrations in leaves on shrubs subjected to water stress for 5 weeks did not recover following re-irrigation, unlike those subjected to water stress for 3 weeks. The leaves on shrubs subjected to water stress for 5 weeks maintained lower levels of chlorophyll during reirrigation. The net rate of CO₂ assimilation (A_n) decreased significantly with an increasing duration of water stress. Reirrigation reversed the decrease in A_n in leaves on shrubs subjected to water stress for 3 weeks. Stomatal conductance (g_s) exhibited a similar pattern to A_n. The actual quantum yield of photosystem II (Φ_PSII) and the electron transport rate (ETR) also decreased significantly with an increasing duration of water stress, although the F_v/F_m ratio was not affected. Φ_PPSII and ETR values in the leaves on shrubs subjected to water stress for 5 weeks did not recover after reirrigation, unlike those subjected to water stress for 3 weeks. Non-photochemical quenching increased with an increasing duration of water stress, but subsequent re-irrigation did not reverse the increase. These results indicate that the timing of re-irrigation of water-stressed ‘Bluecrop’ highbush blueberry is critical in order to maintain their photosynthetic capacity. Among the photosynthetic characteristics measured, Φ_PSII and ETR could be used as sensitive indicators to assess the physiological status of leaves of ‘Bluecrop’ highbush blueberry growing under water stress conditions.     

Sana Physiological responses of two grapevine (Vitis vinifera L.) cultivars to CycocelTM treatment during drought

The plant growth regulator Cycocel^TM [(2-chloroethyl)trimethylammonium chloride] can be used to produce drought tolerance in grapevine (Vitis vinifera L.) due to a reduction in the ratio between vegetative growth and fruit production. To evaluate the physiological responses of two grapevine cultivars to drought and Cycocel^TM treatment, a factorial experiment was conducted in a greenhouse. The factors included irrigation frequency (at 5-, 10-, or 15-day intervals corresponding to no, mild, or severe drought stress), Cycocel^TM concentration (0, 500, or 1000 mg l^–1), and cultivar (‘Rasheh’ or ‘Bidane-Sefid’). Stomatal conductance (g_s) the net rate of CO₂ assimilation (A_net), the rate of transpiration (T_r), and chlorophyll a and b concentrations decreased in plants exposed to mild or severe water-deficit stress, whereas carotenoid, proline, and total soluble sugar concentrations increased compared to plants with no drought stress. The relative water content (RWC) of leaves declined only under severe drought stress. A reduction in intercellular CO₂ concentrations (C_i) occurred under mild drought stress; however, under severe drought stress, C_i values increased. Under mild drought stress, the reduction in the net rate of photosynthesis was related to stomatal closure, whereas under severe drought stress, non-stomatal factors were dominant. Water-use efficiency (WUE) improved under mild drought stress relative to non-stressed plants, but under severe drought, it declined. Foliar applications of Cycocel^TM resulted in increased A_net, g_s, T_r, and WUE values, as well as proline and soluble sugar concentrations. ‘Rasheh’ was more tolerant to drought stress than was ‘Bidane-Sefid’. Foliar applications of Cycocel^TM, particularly at 1000 mg l^–1, mitigated the negative effects of drought stress by increasing A_net, WUE, RWC, compatible solute concentrations, such as proline, soluble sugar, and chlorophyll a and b concentrations.     

In vitro induced tetraploid of Dendranthema nankingense (Nakai) Tzvel. shows an improved level of abiotic stress tolerance

The tetraploid of Dendranthema nankingense (Nakai) Tzvel. was induced by the colchicine treatment using nodal segments. Ploidy level was determined by an analysis of flow cytometry and chromosome counting. The morphological characteristics such as the stomata, leaves, flowers and pollen grains of the tetraploid were significantly larger than those of the diploid. The tolerance responses of the diploid and tetraploid were compared under the imposition of heat, cold, drought and salinity stress. Semi-lethal temperatures suggest that cold tolerance is improved by polyploidization, but the heat tolerance is reduced. Under drought and salt stress, the activity of peroxidase (POD) and relative water content (RWC) in the tetraploid were higher than those in the diploid. Accordingly its malondialdehyde (MDA) content maintained a lower level. The content of chlorophyll (a + b) in the tetraploid was higher, and decrease of its content was postponed in tetraploid compared with the diploid under salt stress. It suggested that polyploidization could alleviate oxidative stress, maintain good water balance and higher chlorophyll (a + b) content, thereby enhanced the drought and salt tolerance in colchicine induced tetraploid.     

Response to water stress of some oleander cultivars suitable for pot plant production

Oleander (Nerium oleander L.) is an evergreen shrub of great ornamental interest which, in recent times, has been increasingly used as a flowering pot plant. Plants grown in pots undergo more frequent water stress conditions than those grown in the soil, due to the limited volume of substrate available for the roots. Oleander is a species adaptable to dry conditions and able to survive long periods of drought. It is well known that under water stress conditions all plants reduce photosynthetic activity, resulting in reduced plant growth. In case of severe water stress, leaves undergo strong wilting and senescence resulting in the loss of ornamental value. In the present work, a study was conducted to evaluate the ecophysiological response to water stress in four oleander cultivars previously recognised (on the basis of traits such as size, habit, earliness, abundance and duration of flowering, aptitude for cutting propagation and rapidity of growth) as suitable for pot plant production. Our data confirm the high drought tolerance of oleander. In the studied cultivars, plants submitted to water stress showed only minor variations in leaf gas exchange parameters [transpiration (E), stomatal conductance (g_s) and CO₂ net assimilation (A)] for at least 10 days without a change in leaf water content [assessed as relative water content (RWC)] for 22 days from the beginning of the stress treatment. During this period, non-irrigated plants maintained the same water status as control plants and were visually undistinguishable from them. Moreover, plants survived without water supply for one month. Following the supply of water again, they were able to restore RWC, gas exchange parameters and instantaneous water use efficiency [A/E ratio (WUE_inst)] to the values of control plants. Furthermore, if at the end of the stress period plants appeared withered and were pale green in colour, they regained their normal appearance after they were irrigated again. Although the four studied cultivars showed some minor differences in leaf gas exchange parameters and in the manner in which the latter parameters changed after irrigation was stopped, the response to water stress was essentially the same. Therefore, as far as drought tolerance is concerned, all these cultivars have a good aptitude for use as flowering pot plants.     

茶树低温胁迫转录因子CsICE的亚细胞定位及表达分析

 以茶树[Camellia sinensis（L.）O. Kuntze]品种‘迎霜’为试验材料,采用PCR技术扩增出与茶树低温胁迫相关转录因子CsICE的开放阅读框,长度783 bp。采用荧光定量PCR分析CsICE的表达量,结果表明CsICE在根、茎、叶、花和果中都有表达,在叶中表达量最高,是根、茎、果中表达量的4倍;在500 μmol ? L^-1 ABA处理下,幼叶中CsICE表达峰值是对照的40倍,在4 ℃低温处理下,最高值是对照的20倍,在10%聚乙二醇6000处理的干旱条件下表达量最高值只有对照的5倍。亚细胞定位结果显示CsICE定位在细胞核上。     

CaCl_2对NaCl胁迫下酸枣苗AsA-GSH循环的影响

以酸枣(Ziziphus acidojujuba C.Y.Cheng et M.J.Liu)水培实生幼苗为试材,研究了CaCl_2对NaCl胁迫下其根、茎、叶中抗坏血酸—谷胱甘肽循环(AsA-GSH)的影响。结果表明:与对照(日本园试配方营养液)相比,NaCl处理下根和叶中AsA含量升高,MDHAR活性升高,APX活性下降;根、茎、叶中GSH含量升高。NaCl+CaCl_2处理与NaC处理相比,根和茎中AsA含量进一步升高,叶中AsA含量下降,叶中APX下降,根中MDHAR活性进一步升高,茎和叶中DHAR活性升高;根和叶中GSH含量下降。外源CaCl_2可能通过提高酸枣实生苗根、茎和叶中AsA再生关键酶MDHAR和DHAR活性,促进根和茎中AsA的再生与生物合成,从而提高根、茎清除过量H_2O_2的能力,缓解NaCl胁迫对酸枣幼苗根和茎部的伤害。     

抗旱性不同的苹果属植物水分胁迫下核酸代谢及自由基变化

 水分胁迫引起苹果属植物中抗旱性较弱的平邑甜茶和抗旱性较强的新疆野苹果幼苗叶片相对含水量(RWC) 、DNA、RNA 含量下降, O₂^-·、H₂O₂ 含量增高, DNase、RNase 活性上升; 其中DNA 含量下降的幅度小于RNA , DNase 活性上升的幅度小于RNase。抗旱性较强的新疆野苹果在水分胁迫下幼苗叶片的RWC、DNA、RNA 含量下降的幅度以及O₂^-·、H₂O₂含量增高、DNase、RNase 活性上升的幅度均小于抗旱性较弱的平邑甜茶。水分胁迫下平邑甜茶和新疆野苹果幼苗叶片的DNA、RNA 含量与RWC、O₂^-·、H₂O₂ 含量和DNase、RNase 活性间均呈显著相关。     

土壤干旱条件下CaCl_2浸种对萝卜幼苗生理指标的影响

在土壤不同干旱程度下研究了60 mmo.lL-1的CaC l2浸种对萝卜幼苗叶片水分状况、光合作用及呼吸强度的影响。结果表明,CaC l2浸种可以使萝卜幼苗离体叶片的保水能力在自然气干时增强;轻度和重度土壤干旱时,无论是CaC l2浸种还是蒸馏水浸种(对照),萝卜幼苗叶片相对含水量、水势、叶绿素含量、光合速率、蒸腾速率和呼吸强度都减小,但CaC l2浸种的下降幅度均比对照小。     

Evidence of drought-sensitive periods from flowering to maturity on highbush blueberry

The effects of water deficit on highbush blueberry (Vaccinium corymbosum L.) have been little studied. This study, conducted on container-planted shrubs, aimed at determining the influence of water stress on growth, water relations and fruit production of the plant during two consecutive years. Drought periods of approximately 3 weeks, as monitored from the transpiration of control plants, were applied at various phenological stages of plant development, from blossoming to harvest, so as to evaluate their direct effects. Highbush blueberry reacted very quickly to drought by reducing transpiration and stopping stem diameter growth and shoot elongation. Its ability to recover depended on the stress level and the drought implementation dynamics. Under moderate stress (35% less transpiration than a well-watered shrub) the recovery potential was almost complete. In all cases water stress during fruit growth and ripening strongly influenced yield by reducing the mean fruit weight and size. Drought after-effects, as assessed from one season to the other, were small, except when stress occurred during flower induction. In that case the number of flowers was reduced in the following year as well as the number of fruits, although fruit size was greater resulting in only a slight reduction in yield. In all cases, photosynthetic performance during the following year appeared not to alter, but water stress imposed to obtain large fruit cannot replace pruning without jeopardising the shrub architecture.     

高丛蓝莓对干旱胁迫的生理响应及其抗旱性综合评价

 为研究高丛蓝莓对干旱胁迫的生理响应并筛选具有较强抗旱能力的品种,同时确定基于生理指标的隶属函数分析对于筛选蓝莓抗干旱品种的可行性,分别以北高丛蓝莓中的‘布里吉塔’、‘斯巴坦’以及南高丛蓝莓中的‘夏普蓝’、‘奥尼尔’4 个品种为材料,在人工控水模拟水分胁迫条件下,测定叶片相对含水量、电导率、丙二醛含量、H₂O₂ 含量与　产生速率、F_v/F_m 以及叶片气孔特性等生理生化指标的变化,并采用隶属函数法进行综合评价。结果表明,随着干旱胁迫程度的加深,4 个蓝莓品种叶片的相对含水量、Fv/Fm、气孔开放率和气孔大小均呈下降趋势,而相对电导率、丙二醛含量、H₂O₂ 含量与产生速率以及气孔密度则呈上升趋势。4 个高丛蓝莓品种的抗旱性强弱顺序为奥尼尔 > 夏普蓝 > 布里吉塔 > 斯巴坦,大田干旱试验验证了基于生理指标的隶属函数分析对于蓝莓抗旱品种的筛选的准确性及可靠性。     

三种暖季型草坪草在干旱条件下脯氨酸含量和抗氧化酶活性的变化

 测定了土壤干旱条件下矮生狗牙根、沟叶结缕草和地毯草相对含水量和相对电导率的变化。结果表明，矮生狗牙根的耐旱性较强，沟叶结缕草次之，而地毯草对干旱敏感。干旱胁迫下，地毯草积累脯氨酸最多，沟叶结缕草次之，矮生狗牙根最少，草坪草的脯氨酸的积累与耐旱性呈负相关。地毯草超氧化物歧化酶(SOD)活性持续升高，过氧化氢酶(CAT)活性在短暂升高后降低，过氧化物酶(POD)活性降低；矮生狗牙根和沟叶结缕草的SOD活性先升后降，CAT和POD活性则维持较高水平，表明草坪草的耐旱性与抗氧化酶活性有关。     

Molecular characterisation and expression analysis of a sHSP gene involved in heat shock treatment-induced chilling tolerance in highbush blueberry fruit

Plant small heat shock proteins (sHSPs) are abundant and diverse proteins involved in various stresses. In this study, a small heat shock protein gene (VcHSP17.7) encoding a 160-amino acid protein was isolated from the highbush blueberry (Vaccinium corymbosum L.). Sequence alignment and phylogenetic analysis indicated that the VcHSP17.7 is a member of the plant cytosolic class II sHSPs; this finding was supported by the subcellular localisation pattern of a VcHSP17.7-YFP fusion protein. Quantitative real-time PCR (qRT-PCR) showed that the VcHSP17.7 gene was expressed in all analysed tissues. Heat shock experiments were conducted using mature blueberry fruit. The anthocyanin content was lower in heat shock-treated fruit than in control fruit after 14 days of storage at 2°C. The malondialdehyde (MDA) content was lower in heat shock-treated fruit than in control fruit during subsequent low-temperature storage, which indicates that heat shock treatment may have alleviated chilling injury (CI) and enhanced the chilling tolerance of the blueberry fruit. Meanwhile, the expression of VcHSP17.7 in blueberry fruit was induced by heat shock treatment and gradually increased during subsequent low-temperature storage. These results suggest that VcHSP17.7 might be involved in the chilling tolerance of blueberry fruit caused by heat shock treatment.     

Water stress induces in pear leaves the rise of betaine level that is associated with drought tolerance in pear

Summary

Mild water stress signi®cantly induced the accumulation of betaine in the leaves of young pear plants (Pyrus bretschneideri Rehd. `Suly'). The betaine induced by water stress in mature leaves was maintained for about two weeks of re-watering after water stress, and gradually declined two weeks after the relief of water stress, dropping to the normal level one month later. Exogenously foliar-applied betaine slightly improved the shoot growth and the ability of the leaves to resist dehydration, alleviated wilting of leaves under water stress, and promoted recovery of leaves from wilting after re-watering. These ®ndings indicate that the accumulated betaine induced by water stress was associated with drought tolerance in pear.     

高锌胁迫对八棱海棠幼苗锌积累、分配与9个MTP基因表达的影响

为了研究高锌(Zn)胁迫下苹果砧木八棱海棠(Malus robusta Rehd.)Zn积累、分配及分子响应机制,以其8叶龄实生苗为试验材料,采用溶液培养法进行不同浓度(4、25、50、100μmol·L~(-1))Zn处理。试验结果表明:过量Zn胁迫严重影响植株生长及Zn的积累与分配,随着Zn处理浓度的升高,八棱海棠幼苗根系中Zn含量显著高于茎和叶;富集系数(BAF)的增大和转运系数(TF)的减小都说明Zn在根系中大量积累,以减轻对地上部的伤害。根据对9个MTP基因的相对表达量以及相关性分析,认为在高Zn胁迫下MTP基因的表达与植株内Zn含量有密切联系。MTP1在茎、叶中显著表达,具有组织特异性。MTPC2-like的相对表达量在4μmol·L~(-1) Zn处理下与根、茎、叶的Zn含量显著正相关,而在较高浓度Zn胁迫下相关性减弱,这说明MTP基因的表达具有浓度效应。