Mulberry Leaf Metabolism under High Temperature Stress

Effects of high temperature on the activity of photosynthetic enzymes and leaf proteins were studied in mulberry (Morus alba L. cv. BC2-59). A series of experiments were conducted at regular intervals (120, 240 and 360 min) to characterize changes in activities of ribulose-1,5-bisphosphate carboxylase (RuBPC) and sucrose phosphate synthase (SPS), photosystem 2 (PS 2) activity, chlorophyll (Chl), carotenoid (Car), starch, sucrose (Suc), amino acid, free proline, protein and nucleic acid contents in leaves under high temperature (40 °C) treatments. High temperature markedly reduced the activities of RuBPC and SPS in leaf extracts. Chl content and PS 2 activity in isolated chloroplasts were also affected by high temperature, particularly over 360 min treatment. Increased leaf temperature affected sugar metabolism through reductions in leaf starch content and sucrose-starch balance. While total soluble protein content decreased under heat, total amino acid content increased. Proline accumulation (1.5-fold) was noticed in high temperature-stressed leaves. A reduction in the contents of foliar nitrogen and nucleic acids (DNA and RNA) was also noticed. SDS-PAGE protein profile showed few additional proteins (68 and 85 kDa) in mulberry plants under heat stress compared to control plants. Our results clearly suggest that mulberry plants are very sensitive to high temperature with particular reference to the photosynthetic carbon metabolism.     

Water stress effects on photosynthesis in different mulberry cultivars

The effect of water stress on photosynthesis was determined in five mulberry cultivars (Morus alba L. cv. K-2, MR-2, BC2-59, S-13 and TR-10). Drought was imposed by withholding water and the plants were maintained at different water potentials ranging from 0.5 -MPa to 2.0 -MPa. Photosynthetic rates, activities of ribulose-1,5-bisphosphate carboxylase and sucrose phosphate synthase, photosystem II activity and chlorophyll content were used as key parameters to assess photosynthetic performance. There was a marked variation in the photosynthetic rates and ribulose-1,5-bisphosphate carboxylase activity among the five mulberry cultivars subjected to water stress. Photosystem II (PSII) and sucrose phosphate synthase activities were also severely reduced as measured by drought conditions. Of the five mulberry cultivars, S-13 and BC2-59 showed higher photosynthetic rates, ribulose-1,5-bisphosphate carboxylase activity, high sucrose phosphate synthase activity and photochemical efficiency of PSII compared to the other varieties.     

Sucrose Metabolism at Three Leaf Development Stages in Bean Plants

Sucrose metabolism was studied at three leaf development stages in two Phaseolus vulgaris L. cultivars, Tacarigua and Montalban. The changes of enzyme activities involved in sucrose metabolism at the leaf development stages were: (1) Sink (9-11 % full leaf expansion, FLE): low total sucrose phosphate synthase (SPS) activity, and higher acid invertase (AI) activity accompanied by low sucrose synthase (SuSy) synthetic and sucrolytic activities. (2) Sink to source transition (40-47 % FLE): increase in total SPS and SuSy activities, decrease in AI activity. (3) Source (96-97 % FLE): high total SPS activity, increased SuSy activities, decreased AI activity. The hexose/sucrose ratio decreased from sink to source leaves in both bean cultivars. The neutral invertase activity was lower than that of AI; it showed an insignificant decrease during the sink-source transition. This revised version was published online in June 2006 with corrections to the Cover Date.     

Photosynthesis and the Enzymes of Photosynthetic Carbon Reduction Cycle in Mulberry During Water Stress and Recovery

Net photosynthetic rate (P _N), stomatal conductance (g _S), transpiration rate (E), intercellular CO₂ concentration (C _i), leaf water potential (_w), leaf area, chlorophyll (Chl) content, and the activities of photosynthetic carbon reduction cycle (PCR) enzymes in two mulberry (Morus alba L.) cultivars (drought tolerant Anantha and drought sensitive M-5) were studied during water stress and recovery. During water stress, P _N, g _S, and E declined whereas C _i increased. P _N, g _S, and E were less affected in Anantha than in M-5, which indicates tolerance nature of Anantha over M-5. Activities of ribulose-5-phosphate kinase, NAD- and NADP-glyceraldehyde-3-phosphate dehydrogenases, and 3-phosphoglycerate kinase decreased with increasing stress in both the cultivars. The enzyme activities less affected in tolerant (Anantha) than in sensitive cultivar (M-5) were restored after re-watering to almost initial values in both the cultivars. Re-watering of the plants led to an almost complete recovery of P _N, E, and g _S, indicating that a short-term stress brings about reversible effect in these two cultivars of mulberry.     

Variation in Photosynthetic Rates and Biomass Productivity among Four Mulberry Cultivars

Among four mulberry (Morus alba L.) cultivars (K-2, MR-2, BC2-59, and S-13), highest net photosynthetic rate (P _N) was observed in BC2-59 while the lowest rates were recorded with K-2. Significant differences among the four cultivars were found in leaf area, biomass production, activities of ribulose-1,5-bisphosphate carboxylase and sucrose phosphate synthase, and glucose and sucrose contents. The P _N and the activities of photosynthetic enzymes in the four cultivars were significantly correlated with the growth and biomass production measured as leaf yield, total shoot mass, and aerial plant biomass.     

Development of Chloroplast in Dark-Grown Wheat Seedlings Irradiated at Elevated Temperature

Elevated temperature inhibited the accumulation of chlorophyll and photosynthetic proteins, and the development of photochemical activity, however, carotenoids continued to accumulate. Signal transduction pathway involved in protochlorophyllide oxidoreductase was unaffected by elevated temperature of 38°C. Two-dimensional gel electrophoresis of stroma proteins showed similar patterns in the dark-grown seedlings and seedlings irradiated at elevated temperature, although some low molecular mass proteins accumulated at 38°C. In contrast, seedlings irradiated at 25°C showed complex pattern of proteins. Hence the development of chloroplast and its associated functions during irradiation of etiolated seedlings are inhibited by elevated temperature.     

Effect of Salinity on Photosynthesis and Biochemical Characteristics in Mulberry Genotypes

Mulberry genotypes were subjected to salinity (0–12 mS cm^–1) in pot culture experiment. Chlorophyll and total carotenoid contents were reduced considerably by salinity. At low salinity, photosynthetic CO₂ uptake increased over the control, but it decreased at higher salinity. Contents of soluble proteins, free amino acids, soluble sugars, sucrose, starch, and phenols increased at salinity of 1–2 mS cm^–1 and decreased at higher salinity (8–12 mS cm^–1). Glycine betaine accumulated more than proline, the maximum accumulation of both was at salinity of 2–4 mS cm^–1. Among the genotypes studied, BC2-59 followed by S-30 showed better salinity tolerance than M-5.     

垂丝海棠应对盐碱复合胁迫的生理响应

盐碱复合胁迫是影响植物生长的主要环境因子。为了探究垂丝海棠（Malus halliana Koehen）响应盐碱复合胁迫的生理特性,利用主成分分析法明确耐盐碱阈值,以2年生垂丝海棠苗为试材,将NaCl与NaHCO₃按1：1的比例混合,通过盆栽浇灌Hogland营养液的方法,模拟5种不同浓度（0、50、100、150、200 mmol/L）盐碱复合胁迫对其光合色素、光合参数、叶绿素荧光参数及渗透调节物质的影响,并对其相关指标进行相关性和主成分分析。结果表明,随着胁迫强度的增大,叶绿素（Chl T）合成受阻,净光合速率（P_n）、气孔导度（G_s）、蒸腾速率（T_r）、胞间CO₂浓度（C_i）、叶片含水量（WC）下降,最大光化学效率（F_v/F_m）、表观光合电子传递速率（ETR）、PSⅡ实际化学效率（φPSⅡ）、光化学猝灭系数（qP）被抑制;类胡萝卜素（Car）含量、水分利用效率（WUE）、电解质外渗率、非调节性能量耗散（Y（NO））升高;胁迫40 d时,初始荧光（F₀）显著升高,而调节性能量耗散Y（NPQ）、非光化学荧光淬灭系数（qN）急剧降低;脯氨酸、可溶性糖、有机酸显著积累,并在100 mmol/L浓度下达到峰值。植株的P_n与G_s、C_i、ETR、φPSⅡ呈极显著正相关,与T_r、WC、qP、Chl T呈显著正相关,与电导率、脯氨酸、Y（NO）呈显著负相关。因此,盐碱复合胁迫下,垂丝海棠叶片主要通过降低G_s、C_i、T_r、WC,提高WUE、大量积累渗透调节物质、启动热耗散机制来保持光合系统伤害与修复的动态平衡。根据主成分分析法,浓度100 mmol/L为耐盐碱阈值,同时G_s、C_i、T_r、Chl T、ETR、φPSⅡ、Y（NO）、WC、qP、电解质渗透率、脯氨酸可作为评价垂丝海棠耐盐碱能力的有效指标。     

Photosynthetic Characteristics in Mulberry during Water Stress and Rewatering

Three-month-old plants of mulberry (Morus alba L. cv. Kanva-2) were subjected to a drought stress by withholding water supply. As the leaf water potential (W) was dropping progressively with the severity of treatment and increasing stress duration, the values of leaf area, dry mass accumulation, total chlorophyll (Chl) content, net photosynthetic rate (PN), stomatal conductance (gs), and transpiration rate (E) were declined. The photosystem 2 (PS2) photochemical efficiency significantly decreased only at a severe stress treatment. The intercellular CO2 concentration (Ci) remained unaltered during a mild stress, yet it increased under moderate and severe stresses. The Ci/gs ratio reflected the mesophyll efficiency during water stress. Rewatering of the plants led to an almost complete recovery of PN, E, and gs, indicating that a short-term stress brings about reversible effects only.     

The Pattern of Changes in Photosynthetic Apparatus in Response to Cold Acclimation and De-acclimation in Two Contrasting Cultivars of Oilseed Rape

In spring and winter cultivars of oilseed rape (Brassica napus var. oleifera), acclimation of photosynthetic apparatus to cold was connected with the increase in activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) and sucrose-phosphate synthase (SPS). Conversely, cold de-acclimation entailed the decline of RuBPCO and SPS activities. The rate of this photosynthetic de-acclimation might depend on day temperature. On the other hand, temperature rise during de-acclimation (identical during the day and night) resulted in the improvement of photosynthetic activity measured by means of chlorophyll fluorescence. An increase in SPS activity (and even transitory increase in RuBPCO activity) was observed when the elongation growth rate (EGR) accelerated during de-acclimation. Throughout re-acclimation, plants with high EGR were unable to maintain or recover higher photosynthetic capacity, despite the fact that SPS activity remained high or even increased during re-acclimation.     

Response of Tea Plants to Water Stress

Two-year-old potted plants of six Camellia sinensis cultivars (TV-18, TV-26, UPASI-3, UPASI-26, T-78 and HV-39) were subjected to water stress for 4, 8 and 12 d. Relative water content (RWC) of leaves of all cultivars declined with water stress, but in the two drought tolerant cultivars (UPASI-3 and UPASI-26), higher RWC were maintained in comparison to the others. Phenol content and activities of phenylalanineammonialyase, polyphenoloxidase and peroxidase initially increased, but decreased during extended drought. Chlorophyll contents decreased, whereas proline contents increased during water stress. SDS-PAGE analysis of proteins revealed increased accumulation of proteins of intermediate molecular masses (42 – 44 kDa) and low molecular masses (14 – 26 kDa). After 12 d of water stress, most of these proteins disappeared in T-78 and HV-39, but in the other cultivars they were still detectable.     

桑树磷脂酶MaPLA1-2D亚型4个基因的克隆与胁迫应答分析

磷脂酶(phospholipase)是一类在植物生长发育和胁迫应答中起重要调控作用的磷脂水解酶,也是一类重要的信号转导酶。而磷脂酶A1(PLA1)在植物应答生物胁迫和非生物胁迫中的功能研究鲜见报道。研究从桑树(Morus alba L.)中克隆了磷脂酶PLA1的1个亚型MaPLA1-2D基因,对其进行了序列分析、组织表达、胁迫诱导表达和蛋白亚细胞定位分析。结果表明,桑树PLA1-2D亚型基因包括4个成员,命名为MaPLA1-2D.1~MaPLA1-2D.4。4个基因在桑树根和叶中高水平表达,蛋白亚细胞定位在叶绿体。序列和进化分析表明MaPLA1-2D基因4个成员与拟南芥AtDAD1基因的保守结构域序列具有较高相似度且进化关系紧密。MaPLA1-2D基因4个成员的启动子含有多种胁迫应答顺式元件和激素响应元件;胁迫诱导表达模式分析表明MaPLA1-2D基因表达受干旱和脱落酸处理显著诱导。以上结果说明,MaPLA1-2D基因与拟南芥DAD同源,可能在桑树非生物胁迫应答中发挥重要功能。     

Effect of Water Stress on Photosynthesis in Two Mulberry Genotypes with Different drought Tolerance

Three-month-old mulberry (Morus alba L.) cultivars (drought tolerant S13 and drought sensitive S54) were subjected to water stress for 15 d. Water stress decreased the leaf water potential, net photosynthetic rate (PN), and stomatal conductance (gs) in both the cultivars. However, the magnitude of decline was comparatively greater in the sensitive cultivar (S54). Intercellular CO2 concentration (Ci) was unaltered during mild stress, but significantly increased at severe stress in both cultivars. The photosystem 2 activity significantly declined only at a severe stress in both cultivars. The Ci/gs ratio representing the mesophyll efficiency was greater in the tolerant cultivar S13. Involvement of stomatal and/or non-stomatal components in declining PN depended on the severity and duration of stress. However, the degree of non-stomatal limitations was relatively less in the drought tolerant cultivar.     

苹果中磷酸蔗糖合酶家族基因的表达特性及其与蔗糖含量的关系

磷酸蔗糖合酶(sucrose phosphate synthase,SPS)是植物中蔗糖合成的主要限速酶,影响植物的生长发育和果实中蔗糖的含量。为探明苹果中SPS基因家族特性及其在蔗糖合成中的作用,该研究从苹果基因组中分离了MdSPS家族基因,分析了它们的进化关系以及mRNA表达特性与酶活性和蔗糖含量的关系。结果显示:(1)在苹果基因组中有8个SPS家族基因表达,它们分别属于双子叶植物的3个SPS亚家族。(2)荧光定量PCR分析显示,苹果C类的MdSPS6基因和A类的MdSPS1a/b基因是苹果中表达丰度最高的SPS基因成员,其中MdSPS6在苹果成熟果中表达丰度最高,其次是成熟叶片,而MdSPS1a/b在不积累蔗糖的幼果中表达丰度最高。(3)在果实发育过程中,除MdSPS1a/b之外,其它5个苹果MdSPS家族基因均随果实的生长表达丰度增加,与SPS活性和蔗糖含量明显呈正相关关系。研究表明,C类家族MdSPS6是苹果果实发育后期和叶片中蔗糖合成的主要SPS基因。     

干旱胁迫对宁夏枸杞叶片蔗糖代谢及光合特性的影响

以宁夏枸杞品种‘宁杞1号’为材料,采用防雨棚内盆栽控水法研究了轻度干旱(LD)、中度干旱(MD)、重度干旱(SD)和正常灌水(CK)4种土壤水分条件下枸杞叶片糖分含量、蔗糖代谢酶活性和光合参数的响应规律,为宁夏枸杞在干旱地区高产栽培提供参考依据。结果显示:(1)干旱胁迫降低了枸杞青果期叶片蔗糖和淀粉含量,轻度干旱胁迫可提高枸杞色变期叶片果糖和蔗糖含量,而成熟期叶片淀粉含量则随着干旱加重而升高。(2)干旱胁迫降低了青果期枸杞叶片中性转化酶(NI)、蔗糖磷酸合成酶(SPS)和蔗糖合成酶(SS)的活性,降低了色变期和成熟期叶片酸性转化酶(AI)和SS的活性,但提高了色变期和成熟期叶片NI的活性。(3)随着干旱胁迫的加剧,叶片净光合速率(Pn)、蒸腾速率(Tr)和气孔导度(Gs)呈降低趋势,瞬时水分利用率(WUE)呈现:LD>CK>SD>MD的变化趋势。研究认为,轻度干旱胁迫能促进宁夏枸杞成熟期叶片蔗糖积累和水分利用率提高,有利于更多光合同化物输送到果实中。     

Water Stress Induced Changes in Anatomy of Tomato Leaf Epidermes

Anatomical changes of leaf epidermes of tomato plants (Lycopersicon esculentum Mill. cv. INCA 9) submitted to water stress in the preflowering stage were studied. 20 d after germination, plants were subjected to three treatments: 1) 100 % of evapotranspired water was applied every day, 2) from 100 up to 10 % of evapotranspired water was applied every day, and 3) water supply was completely suppressed. Trichome density was similar in apical, middle and basal zones, and adaxial and abaxial leaf surfaces. Stomatal density and length, and epidermal cell length and width had similar values on the same leaf surface, but the values were higher on the abaxial than on the adaxial leaf surface. The water deficit had little effect on number of trichomes, length and width of epidermal cells and length of stomata, and decreased the stomatal density especially on adaxial surface.     

Changes in the activities of carbon metabolizing enzymes with pod development in lentil (<Emphasis Type="Italic">Lens culinaris</Emphasis> L.)

Activities of some key enzymes of carbon metabolism sucrose synthase, acid and alkaline invertase, phosphoenol pyruvate carboxylase, malic enzyme and isocitrate dehydrogenase were investigated in relation to the carbohydrate status in lentil pods. Sucrose remained the dominant soluble sugar in the pod wall and seed, with hexoses (glucose and fructose) present at significantly lower levels. Sucrose synthase is the predominant sucrolytic enzyme in the developing seeds of lentil (Lens culinaris L.). Acid invertase was associated with pod elongation and showed little activity in seeds. Sucrose breakdown was dominated by alkaline invertase during the development of podwall, while both the sucrose synthase and alkaline invertase were active in the branch of inflorescence. A substantial increase of sucrolytic enzymes was observed at the time of maximum seed filling stage (10–20 DAF) in lentil seed. The pattern of activity of sucrose synthase highly paralleled the phase of rapid seed filling and therefore, can be correlated with seed sink strength. It seems likely that the fruiting structures of lentil utilize phosphoenol pyruvate carboxylase for recapturing respired carbon dioxide. Higher activities of isocitrate dehydrogenase and malic enzyme in the seed at the time of rapid seed filling could be effectively linked to the deposition of protein reserves.     

NaCl胁迫对祁连山野生黄瑞香叶片光合叶绿素荧光特性的影响

以4片真叶黄瑞香幼苗为材料,设置不同浓度(0、50、100、150、200mmol·L~(-1))NaCl胁迫处理,采用温室砂培实验系统考察了其幼苗叶绿素含量、叶绿素荧光参数及气体交换参数等光合生理指标的变化。结果表明:(1)在正常环境条件下(对照),黄瑞香叶片净光合速率(P_n)、气孔导度(G_s)的日变化曲线呈双峰型,蒸腾速率(T_r)日变化曲线呈单峰型;较高浓度(100mmol·L~(-1))NaCl胁迫改变了黄瑞香叶片光合特性日变化曲线,导致其P_n、T_r、G_s日变化曲线整体下降,而胞间CO_2浓度(Ci)日变化曲线整体上升。(2)低浓度(50mmol·L~(-1))NaCl胁迫对黄瑞香叶片叶绿素含量及其比值无显著影响,但较高浓度(100mmol·L~(-1))NaCl胁迫则使叶绿素含量显著下降,其比值下降则较平缓。(3)较高浓度(100mmol·L~(-1))NaCl胁迫使得黄瑞香叶片最大荧光(F_m)、PSⅡ最大光化学效率(F_v/F_m)、PSⅡ光下最大捕光效率(F_v′/F_m′)、光化学荧光猝灭系数(qP)、PSⅡ实际光化学效率(Φ_(PSⅡ))均显著下降,却使其初始荧光(F_0)和非光化学猝灭(NPQ)显著上升。研究发现,随着盐胁迫浓度的增加,引起黄瑞香光合速率下降的主要原因是非气孔因素;在轻度NaCl胁迫下黄瑞香有较强的忍耐性,而重度NaCl胁迫则显著降低了叶片的光合机构活性,加剧了光抑制程度,从而严重限制了其叶片的光合作用效率。     

Changes of Photosynthetic Characteristics in Relation to Leaf Senescence in Two Maize Hybrids with Different Senescent Appearance

A field experiment was conducted to investigate the changes in chlorophyll (Chl) and nitrogen (N) contents, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) and phosphoenolpyruvate carboxylase (PEPC) contents and PEPC activity, and the photon-saturated net photosynthetic rate (P _Nsat), and their relationships with leaf senescence in two maize hybrids with different senescent appearance. One stay-green (cv. P3845) and one earlier senescent (cv. Hokkou 55) hybrid were used in this study, and we found that Chl and N contents and the P _Nsat in individual leaves of P3845 were greater than those in corresponding leaves of Hokkou 55 at the successive growth stages. In addition, larger contents of RuBPCO and PEPC, and a greater activity of PEPC were observed in P3845. Due to the lower rates of decrease of Chl, RuBPCO, and PEPC amounts per unit of N, and the lower net C translocation rate per unit of N in the stay-green hybrid, leaf senescence was delayed in comparison to the earlier senescent hybrid.     

闽楠叶片响应干旱胁迫的蛋白质组分析

闽楠对土壤水分要求较高,为了提高闽楠造林的成活率,该研究以正常供水为对照,测定了不同干旱胁迫时间(7d、14d)及复水后7d的叶片蛋白质组及生理生化指标变化,以探讨闽楠响应干旱胁迫的分子生理机制。结果表明:(1)不同干旱胁迫处理下闽楠叶片蛋白质双向电泳(2-DE)分析结果共发现51个差异表达蛋白;采用MALDI-TOF/TOF成功鉴定到45个蛋白点;这些鉴定出的差异蛋白与光合作用、碳水化合物和能量代谢、胁迫响应与防御、翻译后修饰、蛋白质转换与分子伴侣功能等生理代谢过程密切相关。(2)检测不同干旱处理时间闽楠叶片膜脂过氧化相关MDA含量,防御相关酶SOD、CAT、POD活性和糖代谢相关酶PFK、AGPase、PK及PDH活性,发现各指标在14d持续干旱胁迫时主要呈下降趋势,且变化均达到极显著水平(P0.01)。研究认为,持续干旱胁迫下,光合作用和植物防御系统以及能量和糖代谢的降低是闽楠不耐干旱的重要生理生化原因,研究结果为今后耐旱闽楠的分子育种提供了理论依据。