Farnesol-like antitranspirant activity and stomatal behaviour in maize and Sorghum lines of differing drought tolerance

Abstract Potted seedlings of four lines of maize and Sorghum of differing drought tolerance were subjected to a single soil drying cycle and were only rewatered when the plants showed the first signs of wilting. Other plants remained well-watered throughout the experimental period. As plant water potentials decreased in the unwatered plants of three of the lines investigated (Sorghum Piper and M35-1, V-4146 and maize Farz 27), endogenous levels of farnesol-like antitranspirants increased. Closure of stomata correlated well with the increase in endogenous antitranspirant. In the fourth line (Sorghum M35-1, V-4184), stomata did not close as the level of plant water stress increased, although leaf diffusion resistance of even the well-watered plants of this line was quite high. In this line, there was no consistent relationship between plant water stress and antitranspirant level or between stomatal behaviour and antitranspirant level. The involvement of farnesol-like antitranspirants in the control of stomatal behaviour in water-stressed plants is discussed.     

Heat shock protein expression in thermotolerant and thermosensitive lines of cotton

The expression of heat shock proteins (HSPs) was compared between genetically characterized heat tolerant and heat sensitive lines of cotton (Gossypium hirsutum andG. barbadense) using electrophoretic analysis ofin vivo labelled proteins. No differences were observed between the two lines with regard to: the temperature at which HSP synthesis was induced (37°C); the temperature at which HSP synthesis was maximal (45°C); the rates of recovery from HSP synthesis; the duration of HSP synthesis; or the major size classes of HSPs expressed in these two lines. Several HSPs were identified on 2D gels which were expressed uniquely in either the tolerant or sensitive cotton line. However, the HSP pattern displayed in a heat tolerant BC-3 individual was that of the heat sensitive parent.Abbreviations HSPs heat shock proteins - IEF isoelecticfocusing     

Complexity and Genetic Variability of Heat-Shock Protein Expression in Isolated Maize Microspores

The expression of heat-shock proteins (HSPs) in isolated maize (Zea mays L.) microspores has been investigated using high-resolution two-dimensional electrophoresis coupled to immunodetection and fluorography of in vivo synthesized proteins. To this end, homogeneous and viable populations of microspores have been purified in sufficient amounts for molecular analysis from plants grown in controlled conditions. Appropriate conditions for thermal stress application have been defined. The analysis revealed that isolated microspores from maize display a classical heat-shock response characterized by the repression of the normal protein synthesis and the expression of a set of HSPs. A high complexity of the response was demonstrated, with numerous different HSPs being resolved in each known major HSP molecular weight class. However, the extent of this heat-shock response is limited in that some of these HSPs do not accumulate at high levels following temperature elevation. Comparative analysis of the heat-shock responses of microspores isolated from five genotypes demonstrated high levels of genetic variability. Furthermore, many HSPs were detected in microspores at control temperature, indicating a possible involvement of these proteins in pollen development at stages close to first pollen mitosis.     

Influence of Temperature Stress on in Vitro Fertilization and Heat Shock Protein Synthesis in Maize (Zea mays L.) Reproductive Tissues

This study was conducted to investigate the response of maize (Zea mays) male and female mature reproductive tissues to temperature stress. We have tested the fertilization abilities of the stressed spikelets and pollen using in vitro pollination-fertilization to determine their respective tolerance to stress. The synthesis of heat shock proteins (HSPs) was also analyzed in male and female tissues using electrophoresis of ³⁵S-labeled proteins and fluorography, to establish a relationship between the physiological and molecular responses. Pollen, spikelets, and pollinated spikelets were exposed to selected temperatures (4, 28, 32, 36, or 40°C) and tested using an in vitro fertilization system. The fertilization rate is highly reduced when pollinated spikelets are exposed to temperatures over 36°C. When pollen and spikelets are exposed separately to temperature stress, the female tissues appear resistant to 4 hours of cold stress (4°C) or heat stress (40°C). Under heat shock conditions, the synthesis of a typical set of HSPs is induced in the female tissues. In contrast, the mature pollen is sensitive to heat stress and is responsible for the failure of fertilization at high temperatures. At the molecular level, no heat shock response is detected in the mature pollen.     

Altered gene expression in plants with constitutive expression of a mitochondrial small heat shock protein suggests the involvement of retrograde regulation in the heat stress response

Heat stress can negatively affect crop productivity. One way in which plants attempt to alleviate the effects of heat stress is to induce the expression of genes encoding heat shock proteins (HSPs), including small HSPs (sHSPs). We produced transgenic lines of Arabidopsis thaliana expressing a transgene encoding a maize mitochondrial sHSP, ZmHSP22. The transgene, under the control of the cauliflower mosaic virus 35S promoter, is constitutively highly expressed in these lines. As demonstrated by confocal immunofluorescence microscopy and analyses of isolated mitochondria, ZmHSP22 is directed to the mitochondria of Arabidopsis and is processed into the mature form. These transgenic lines demonstrated altered expression of nuclear genes encoding the endogenous mitochondrial sHSP, AtHSP23.6, chloroplast localized AtHSP25.3, class I cytosolic AtHSP17.4, cytosolic AtHSP70-1 and chloroplast localized AtHSP70-6, but not cytosolic AtHSP70-15, following exposure to heat stress. This suggests that the expression of HSPs can be affected by heat-induced mitochondrial retrograde regulation. Three-week-old plants from the transgenic Arabidopsis lines expressing ZmHSP22 have increased thermotolerance, as measured by the maintenance of higher leaf mass following successive days with short periods of heat stress.     

In vitro selection and regeneration of cotton resistant to high temperature stress

Summary Cell suspension cultures of cotton (Gossypium hitirsutum L. cv. Coker 312) were exposed to various temperature:time treatments in order to select cell lines resistant to high temperature stress. Cells were exposed to 45°C for 3 h each day until the total accumulated hours of stress were: 0 h, 10 h, 75 h, 100 h, or 105 h (81 h pulsed then 24 h continuous). After the stress treatments, the cells were plated onto embryo development medium and plants were recovered. The embryogenic calli that were recovered were subcultured monthly for 6 months and tested for increased resistance to the temperature:time treatments previously determined to be lethal and to water stress as imposed by PEG. All of the selected cell lines were more resistant to both types of stress than the control cell lines. Leaf tissue of stress selected (Ro) formed and maintained callus growth when incubated at 38°C; whereas, tissue excised from nonselected controls rarely formed callus and calli which did form quickly became necrotic. These cells and plants will provide a tool for determining the mechanisms involved in resistance to high temperature stress.     

Root cortical aerenchyma improves the drought tolerance of maize (Zea mays L.)

Root cortical aerenchyma (RCA) reduces root respiration in maize by converting living cortical tissue to air volume. We hypothesized that RCA increases drought tolerance by reducing root metabolic costs, permitting greater root growth and water acquisition from drying soil. To test this hypothesis, recombinant inbred lines with high and low RCA were observed under water stress in the field and in soil mesocosms in a greenhouse. In the field, lines with high RCA had 30% more shoot biomass at flowering compared with lines with low RCA under water stress. Root length density in deep soil was significantly greater in the high RCA lines compared with the low RCA lines. Mid‐day leaf relative water content in the high RCA lines was 10% greater than in the low RCA lines under water stress. The high RCA lines averaged eight times the yield of the low RCA lines under water stress. In mesocosms, high RCA lines had less seminal root respiration, deeper rooting, and greater shoot biomass compared with low RCA lines under water stress. These results support the hypothesis that RCA is beneficial for drought tolerance in maize by reducing the metabolic cost of soil exploration.     

Stomatal Reactions of Two Different Maize Lines to Osmotically Induced Drought Stress

Two maize lines differing in drought resistance were grown at different drought stress induced by polyethylene glycol (PEG 10 000) solutions with osmotic potentials of –0.20, –0.40 and –0.80 MPa in the semipermeable membrane system. During the five days soil water content decreased (from 0.43 to 0.29, 0.25 and 0.23 g cm^–3 for three PEG solutions, respectively) as well as leaf water potentials (_w; from – 0.54 to –0.76, –1.06 and –1.46 MPa). These values were not significantly different between the investigated lines, indicating that a controlled and consistent soil moisture stress was achieved. Soil drying induced an increase in the ABA content of leaves and xylem of both lines and the effects on stomatal conductance were greater in drought susceptible line (B-432) compared to drought resistant line (ZPBL-1304). To test possible difference in stomatal sensitivity to xylem ABA between lines and to assess any ABA vs. _w interaction, roots were fed with 10, 50 and 100 mmol m^–3 ABA solutions in another set of experiments. These results showed that manipulation of xylem ABA affected stomata of both lines similarly. Comparison of stomatal sensitivity to drought-induced and applied ABA demonstrated that drought treatment affected stomata of investigated lines by differentially increasing their sensitivity to xylem ABA, thus confirming an interaction between chemical signalling and hydraulic signalling.     

Heat shock induced proteins in plant cells

Tobacco (Nicotiana tabacum) and soybean (Glycine max) tissue culture cells were exposed to a heat shock and protein synthesis studied by SDS-polyacrylamide gel electrophoresis after labeling with radioactive amino acids. A new pattern of protein synthesis is observed in heat-shocked cells compared to that in control cells. About 12 protein bands, some newly appearing, others synthesized in greatly increased quantities in heat-shock cells, are seen. Several of the heat-shock proteins (HSPs) in both tobacco and soybean are similar in size. One of the HSPs in soybean (76K) shares peptide homology with its presumptive 25°C counterpart, indicating that the synthesis of at least some HSPs may not be due to activation of new genes. The optimum temperature for maximal induction of most HSPs is 39–40°C. Total protein synthesis decreases as heat-shock temperature is increased and is barely detectable at 45°C. The heat-shock response is maintained for a relatively short time in tobacco cells. After 3 hr at 39°C, a decrease is seen in the synthesis of the HSPs, and after 4 hr practically no HSPs are synthesized. After exposure to 39°C for 1 hr, followed by a return of tobacco cells to 26°C, recovery to the control pattern of synthesis requires greater than 6 hours. These results indicate that cells of flowering plants exhibit a heat-shock response similar to that observed in animal cells.     

Role of Abscisic Acid in Thermal Acclimation of Plants

Abscisic acid (ABA) is a stress hormone that confers resistance to abiotic stressors, including drought, salt, cold, and heat. In general, antioxidant capacity and heat shock proteins (HSPs) mainly mediate ABA to enhance thermal acclimation in plants, but sugar metabolism and signaling also play critical roles in this response in the presence of ABA. Indeed, ABA accelerates sugar metabolism and transports more carbohydrates to spikelets under heat stress, which is beneficial to plants surviving under stressful conditions. Few studies have summarized the interactions among sucrose metabolism, signaling, and hormones in plants during heat stress, but this topic will likely attract more attention in the future. This article reviews the antioxidant capacity, HSPs, sugar metabolism, hormone crosstalk, and their interactions involved in ABA-induced heat tolerance in plants. Clarifying the underlying mechanisms will be invaluable for breeding heat-resistant cultivars and for developing new tissue culture techniques that reduce heat damage in plants.     

新疆自然高温环境下玉米自交系开花期耐热性鉴定与评价

在新疆夏季自然高温环境下,以田间玉米空秆率、果穗结实率、相对结实率为主要评价指标,综合籽粒产量与穗部性状,对26份国内外玉米骨干自交系连续2年进行耐热性鉴定与评价,结合新疆当地高温干燥的气候特点,建立一套适宜玉米种质资源大田耐热性的鉴定、评价体系,为中国玉米耐热种质资源的遗传改良和新品种选育提供参考。结果表明:(1)不同基因型玉米自交系空秆率、果穗结穗率、相对结实率和耐热性差异较大,其中GW5F、GW4F、GW7F和PH6WC耐热性最强,在高温胁迫下空秆率最低,果穗结实率与相对结实率最高;其次为PHBA6、新自351等8份自交系,在高温胁迫下籽粒产量及综合表现较好;其余自交系对高温均表现敏感。(2)高温胁迫导致玉米穗部性状严重衰退,其中的穗重、穗行数和行粒数所受影响最大。(3)在年份或材料之间,玉米自交系籽粒产量与其主要农艺性状相关性差异较大,并以果穗重与籽粒产量相关性最高。(4)以相对结实率为主要指标通过层次聚类可将参试材料分为两大群,耐热性最强的GW5F、PH6WC、GW4M聚为第Ⅰ群,其余自交系聚为第Ⅱ群;第Ⅱ群中‘吉63’、Mo17等6份自交系可聚为第1亚群,PHBA6、LH82等6份自交系聚为第2亚群,郑58、新农育6390M等11份自交系聚为第3亚群。综合分析来看,该研究中表现极耐高温的4份自交系均为国外优异种质,可用于玉米耐高温基础研究和遗传改良;表现较强耐热性的8份品系多来自于新疆本地选育的材料,其遗传基础较为广泛,适应当地的高温环境,也是优异的耐热育种资源。     

Analysis of NaCl stress response in Tunisian and reference lines of <Emphasis Type="Italic">Medicago truncatula</Emphasis>

We analyzed the effects of NaCl stress on nine lines of Medicago truncatula, including four lines from southern Tunisia, three from the north, and two references lines. Plants were cultivated in two treatments (0 and 45 mM NaCl) for a period of 45 days. At harvest, we measured seven quantitative traits of the shoot and root growth. The analysis of variance showed that responses of lines to NaCl stress depended on the effects of line, treatment, and their interaction. Treatment had the largest effect. All measured traits showed high broad-sense heritability (H ²) under both treatments. While the leaf area was the less affected trait, the length of stems was the most affected regarding the sensitivity index (SI) parameter. Estimated SI for total plant biomass showed that TN13.11 and TN11.0 were the most contrasting lines with the lowest and the highest values, respectively. Established correlations between measured traits and the hierarchical cluster relationships among lines depended on the treatment effect. The site-of-origin environmental factor that influenced more the response of lines to NaCl stress was the relative humidity.     

Heat shock proteins and effects of heat shock in plants

Soybean seedlings when exposed to a heat shock respond in a manner very similar to that exhibited by cultured cells, and reported earlier [2]. Maximum synthesis of heat shock proteins (HSPs) occurs at 40C. The heat shock response is maintained for a relatively short time under continuous high temperature. After 2.5 hr at 40 C the synthesis of HSPs decreases reaching a very low level by 6 hr. The HSPs synthesized by cultured cells and seedlings are identical and there is a large degree of similarity in HSPs synthesized between the taxonomically widely separated species, soybean and corn. Storage protein synthesis in the developing soybean embryo is not inhibited but is actually stimulated during a heat shock, unlike most other non-HSPs, whose synthesis is greatly reduced. Seedlings respond differently to a gradual increase in temperature than they do a sudden heat shock. There is an upward shift of several degrees in the temperature at which maximum protein synthesis occurs and before it begins to be inhibited. In addition, there appears to be a protection of normal protein synthesis from heat shock inhibition when the temperature increase is gradual. An additional function of the heat shock phenomenon might be the protection of seedlings from death caused by extreme heat stress. The heat shock response appears to have relevance to plants in the field.     

Arbuscular mycorrhizae improves low temperature stress in maize via alterations in host water status and photosynthesis

The effect of arbuscular mycorrhizal (AM) fungus, Glomus etunicatum, on growth, water status, chlorophyll concentration and photosynthesis in maize (Zea mays L.) plants was investigated in pot culture under low temperature stress. The maize plants were placed in a sand and soil mixture at 25°C for 7 weeks, and then subjected to 5°C, 15°C and 25°C for 1 week. Low temperature stress decreased AM root colonization. AM symbiosis stimulated plant growth and had higher root dry weight at all temperature treatments. Mycorrhizal plants had better water status than corresponding non-mycorrhizal plants, and significant differences were found in water conservation (WC) and water use efficiency (WUE) regardless of temperature treatments. AM colonization increased the concentrations of chlorophyll a, chlorophyll b and chlorophyll a + b. The maximal fluorescence (Fm), maximum quantum efficiency of PSII primary photochemistry (Fv/Fm) and potential photochemical efficiency (Fv/Fo) were higher, but primary fluorescence (Fo) was lower in AM plants compared with non-AM plants. AM inoculation notably increased net photosynthetic rate (Pn) and transpiration rate (E) of maize plants. Mycorrhizal plants had higher stomatal conductance (g_s) than non-mycorrhizal plants with significant difference only at 5°C. Intercellular CO₂ concentration (Ci) was lower in mycorrhizal than that in non-mycorrhizal plants, especially under low temperature stress. The results indicated that AM symbiosis protect maize plants against low temperature stress through improving the water status and photosynthetic capacity.     

外源海藻糖对高温胁迫下穗分化期水稻叶片生理特性及产量的影响

该研究以耐热型水稻品种Nagina22和热敏型水稻品种YR343为供试材料,采用盆栽试验,设置喷施清水+常温处理(NT0)、喷施清水+穗分化期高温胁迫(HT0),以及分别喷施5、10、15、20 mmol·L^-1外源海藻糖+高温胁迫(分别记为HT1、HT2、HT3、HT4)共6个处理,分析外源海藻糖对高温胁迫下穗分化期水稻叶片叶绿素含量、光合气体交换参数、抗氧化酶活性、渗透调节物质含量、活性氧含量等生理特性,以及籽粒产量及其构成因素的影响,为水稻抗热栽培和耐热品种的选育提供理论依据。结果表明:(1)在高温胁迫下水稻穗分化期,2个水稻品种叶片的叶绿素含量、光合气体交换参数及渗透调节物质含量均降低,叶片MDA和H₂O₂含量以及■的产生速率均上升,叶片抗氧化酶活性呈先增后降的趋势,最终显示水稻籽粒产量及其构成因素显著下降。(2)喷施外源海藻糖能显著增加高温胁迫下穗分化期水稻的每穗粒数、千粒重和结实率,从而提高籽粒产量,其中弱势粒千粒重和结实率的增幅高于强势粒,外源海藻糖最适喷施浓度为15 mmol·L^-1...