Contribution of the wheat (Triticum aestivum L.) flag leaf to grain yield in response to plant growth regulators

In West-Europe, intensive cereal management uses plant growth regulators (PGRs) especially for wheat. A green-house experiment compared the effects of two PGRs on flag leaf characteristics and yield of winter wheat. Chlormequat chloride + choline chloride (CCC) and chlormequat chloride + choline chloride + imazaquin (CCC+I) were applied to winter wheat at growth stage 5 (Feekes Large scale). CCC and CCC+I significantly increased flag leaf surface area at anthesis. Both treatments also enhanced chlorophyll content of the main stem flag leaf. The grain filling period was extended with PGR application by 2 days. CCC and CCC+I significantly increased net CO₂ assimilation rates during the flag leaf life. No effects of PGR spraying were observed on the pattern of ¹⁴C labelled assimilate distribution. Increased grain yield was due to the increase in average grain weight. The results indicate that PGR treatments increased flag leaf contribution to grain filling. The addition of imazaquin (I) to chlormequat (CCC) improved the effects of CCC.     

Regeneration after Wilting, Growth and Yield of Wheat Plants, as Affected by Two Growth–Retarding Compounds

The effect of spraying with 2–chloroethyltrimethylammonium chloride (CCC) and N-dimethylaminosuccinamic acid (B-995) on wheat plants exposed to various durations of wilting, was studied. The chemicals had little or no effect on dry matter production, grain yield, or water requirement of plants watered regularly or exposed to a short drought period causing one day of wilting. In plants exposed to two drought cycles of 5–6 wilting days each, a very pronounced increase in dry weight and grain production was found in plants treated with the two growth retardants. This effect was due to the increased ability of the treated plants to regenerate new shoots on rewatering after wilting. Longer drought periods of 10–12 wilting days, caused complete desiccation of both treated and untreated plants. The mode of action of the chemicals in increasing the drought tolerance of plants is discussed in relation to their effect on delaying the senescence of detached leaves.     

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。     

Drought and watering-dependent oxidative stress: effect on antioxidant content in Triticum aestivum L. leaves

The purpose of the present work was to evaluate both oxidative stress and the antioxidant response system in leaves from wheat (Triticum aestivum cv. Buck Poncho) subjected sequentially to drought and watering. Drought was imposed by withholding water until soil water potential reached -2.0 MPa and maintained under those conditions for 24 h. DCFDA oxidation by wheat leaves was not significantly affected by drought, but watering led to an approximately 2-fold increase in DCFDA oxidation rate. However, no significant effect either on lipid radical content or on hydroperoxide content was measured after drought and drought followed by watering. Microsomes isolated from leaves exposed to drought, and from leaves exposed to drought followed by watering, generated a significantly higher amount of hydroxyl radical as compared to microsomes isolated for control leaves suggesting a higher production of hydroxyl radical in the cellular water soluble phase, after drought and watering as compared to control values. The content of -tocopherol in wheat leaves was increased 2.4-fold after drought and -carotene content was increased by 2.6-fold after drought. Hydration lowered lipid-soluble antioxidant content to control values. Total thiol content as increased by 70% after drought, and watering did not significantly alter the enhanced values. Drought decreased by 28.5% the content of reduced ascorbic acid. Taken as a whole, active species formed at wheat membranes after exposure to moderate water stress, are efficiently removed upon rehydration by reaction with an increased content of -tocopherol and -carotene. Moreover a co-ordinated response involving glutathione reductase activity, thiols and ascorbic acid is triggered to limit free radical dependent effects.Key words: Antioxidants, lipid radicals, oxygen radicals, water stress, wheat      

Experiments with CCC on wheat: effects of spacing, nitrogen and irrigation

In experiments with spring and winter wheat at Rothamsted and Woburn during 4 years CCC increased yield at close spacing (4 in) (10 cm) more than at usual spacing (8 in) (20 cm), but there was no interaction between spacing and yield. Some experiments tested up to 2·4 cwt/acre (300 kg/ha) N to see whether yields continued to increase with more than usual amounts of N, when CCC prevented lodging. There was no evidence of this. When a short dry spell occurred at ear emergence, yield of spring wheat was increased by 6 cwt/acre (750 kg/ha) by CCC and 10 cwt (1250 kg) by irrigation. CCC probably improves yield in these conditions because the larger root system it causes enables more ear-bearing shoots to survive. CCC increases yield in two ways, either by increasing ears or grain per ear. In an unlodged crop CCC usually makes the grains smaller, but by preventing lodging it can also increase size. Usually CCC decreases the leaf area per shoot. The flag leaf may be smaller, unchanged or larger than those of untreated plants. There was no obvious connexion between flag-leaf area and grain yield; when CCC decreased flag-leaf area duration by 25 %, grain yield was unchanged. The results suggest that using CCC gives a more than even chance of a profitable yield increase.     

The effect of acetylcholine on growth and on growth inhibition by CCC in wheat seedlings

Summary The inhibition of the growth of wheat seedlings by 2-chloroethyltrimethylammonium chloride (CCC) was strongly reduced by root application of acetylcholine (Ach). Ach was applied after uptake of CCC by the roots or by the leaves. Ach also stimulated growth of non-CCC-treated seedlings up to 30% when applied to the roots. Growth stimulation appeared to be dependent on pH of the medium and most effective at pH 4.5 and 6. At pH 7.5 Ach did not promote growth of wheat seedlings.Organization for Applied Scientific Research.—Author's business address: Stichting, Centrum voor Plantenfysiologisch Onderzoek, C.P.O., Postbus 52, Wageningen, The Netherlands.     

Drought,high temperature,and their combination affect ultrastructure of chloroplasts and mitochondria in wheat (Triticum aestivum L.) leaves

Plants experience a number of limiting factors, as drought and heat, which are often coinciding stress factors in natural environment. This study evaluated the changes in mesophyll cell ultrastructure in the leaves of two varieties of winter wheat (Triticum aestivum L.), differing in their drought tolerance, under individual or combined drought and heat treatment. Although the individual stress factors affected leaf ultrastructure, the damaging effect of the combined drought and heat was more pronounced and manifested certain differences between genotypes. Chloroplasts and mitochondria were affected in a variety-specific manner under all adverse treatments. The organelles of the drought-tolerant Katya were better preserved than those in the sensitive variety Sadovo. Leaf ultrastructure can be considered as one of the important characteristics in the evaluation of the drought susceptibility of different wheat varieties.     

Relationship between grain yield, osmotic adjustment and benzoxazinone content in Triticum aestivum L. cultivars

Fifteen wheat genotypes were grown under water deficit to ascertain the role of osmotic adjustment (OA) and the concentration of benzoxazinones in sustaining grain yield. A positive correlation between osmotic adjustment capacity and yield was observed in wheat genotypes cultivated under field conditions. The weight gain of plants exposed to drought was in agreement with the OA values (R(2) = 0.93). However, when wheat plants were infested by cereal aphids, this correlation was not found. The benzoxazinones 2,4-dihydroxy-1,4-benzoxa-zin-3-one (DIBOA) and 2,4-dihydroxy-7-methoxy-1,4 benzoxazin-3-one (DIMBOA) are defensive secondary metabolites present in wheat and others cereals. The content of these compounds varied in wheat genotypes and increased with drought and aphid infestation. A positive correlation between weight gain of irrigated-infested plants and drought-infested plants and the contents of benzoxazinones was observed. These results suggest that plants with better OA capacity and high benzoxazinone content should have better field yields.     

Wheat (Triticum aestivum L.) growth enhancement by Azospirillum sp. under drought stress

Plant growth promoting rhizobacteria (PGPR) can enhance plant growth by alleviating soil stresses. Although previously investigated, some new interesting details are presented regarding the alleviating affects of Azospirillum sp. on wheat growth under drought stress in this research work. We hypothesized that the isolated strains of Azospirillum sp. may alleviate the adverse effects of drought stress on wheat (Triticum aestivum L.) growth. Three different strains of Azospirillum lipoferum (B1, B2 and B3) were used to inoculate wheat seedlings under drought. During the flowering stage the seedlings were subjected to three drought levels with five different time longevity, including control. Pots were water stressed at 80% (S0), 50% (S1) and 25% (S2) of field capacity moisture in a 25 day-period. Soil and plant water properties including water potential and water content, along with their effects on bacterial inoculum and wheat growth, were completely monitored during the experiment. While stress intensity significantly affected bacterial population and wheat growth, stress longevity only affected wheat water potential and water content. Compared to uninoculated treatments strain B3 (fixing and producing the highest amounts of N and auxin, respectively, with P solubilizing and ACC-deaminase activities) increased wheat yield at S1 and S2 by 43 and 109%, respectively. However, strain B2 (producing siderophore) was the most resistant strain under drought stress. The results of this experiment may elucidate the more efficient strains of Azospirillum sp. for wheat inoculation under drought stress and the mechanisms by which they alleviate the stress.     

水分胁迫下不同抗旱性小麦品种叶片转录因子表达差异研究

通过研究不同抗旱性小麦品种中转录因子表达水平的差异,为阐明小麦抗旱机制奠定基础。依据候选基因序列设计PCR引物,以干旱胁迫后0、3、6、9、12和24 h的小麦叶片为实验材料,以26S rRNA为内参,运用荧光定量PCR技术,检测Wdreb2、Wlip19基因在干旱敏感性和干旱耐受性小麦叶片中的相对表达量。定量PCR结果显示：干旱胁迫后,Wdreb2、Wlip19基因在干旱敏感性小麦叶片中的表达明显低于干旱耐受性小麦,在不同品种叶片中的响应时间和表达趋势存在差异。研究认为,Wdreb2、Wlip19基因在不同品种小麦受到干旱胁迫后的表达差异,与该品种小麦的抗旱能力具有一定的相关性。     

Nitrogen Modulates the Effects of Heat,Drought, and Combined Stresses on Photosynthesis,Antioxidant Capacity,Cell Osmoregulation,and Grain Yield in Winter Wheat

Longer and more intense heat and drought stresses will occur in terrestrial ecosystems in the future. Although the effects of individual heat or drought stress on wheat plants have been largely explored, the regulatory effect of nitrogen (N) on winter wheat under heat, drought, and combined stresses and whether N alleviates damage to wheat plants caused by these stresses remain unclear. Therefore, the objective of the present study was to investigate the growth, photosynthesis, antioxidant enzyme and N metabolism-related enzyme activity, cell membrane system, osmoregulatory substance, and yield responses to heat, drought, and combined stresses in wheat plants and to clarify the regulatory effects of N on the growth, physiological and biochemical characteristics, and yield of wheat plants under stress conditions. The results showed that wheat plant exposure to individual heat or drought stress reduced photosynthesis and N metabolism-related enzyme activities and increased antioxidant enzyme activities, electrolyte leakage (EL), and the contents of MDA (malondialdehyde) and O₂^? (superoxide anion). The above parameters showed typical superposition effects under combined stress. Under individual heat or drought stress, wheat plants treated with a medium (N₂) or high (N₃) N supply maintained higher photosynthesis and N metabolism-related enzyme activities than did those treated with a low N supply (N₁). Enhanced heat and drought tolerance in wheat plants under an appropriate N supply may be attributed to improved antioxidant capacity, as exemplified by increased activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), glutathione reductase (GR) and ascorbate peroxidase (APX), and to enhanced osmoregulation capacity, as signified by increased contents of soluble sugar (SS), soluble protein (SP), and proline (Pro). Variable importance in projection (VIP) analysis indicated that efficient SOD, POD, CAT, and GR activities and an increased Pro content had superior potential to alleviate heat, drought, and combined stress stresses in wheat plants, and the improvements in growth and grain yield in wheat plants further confirmed the oxidative stress alleviation and stress tolerance enhancement. However, positive effects of N on wheat growth and grain yield under combined stress were usually observed under a low N supply. These results may facilitate future research on the effects of N fertilizer on the stress resistance of winter wheat.

Graphical Abstract

     

Antioxidant status and physiological responses of wheat (Triticum aestivum L.) to cycocel application and bio fertilizers under water limitation condition

In order to study antioxidant status and physiological responses of wheat to cycocel (CCC) and bio fertilizers application under water limitation condition, a factorial experiment was conducted based on randomized complete block design with three replications in 2015. Treatments included water limitation in three levels [normal irrigation (I₁) as control; moderate water limitation (I₂) or irrigation withholding at 50% of heading stage; severe water limitation (I₃) or irrigation withholding at 50% of booting stage]; four bio fertilizer levels [(no bio fertilizer (F₀), seed inoculation by Azotobacter chrocoocum strain 5 (F₁), Pseudomonas putida strain 186 (F₂), Azotobacter?+?Pseudomonas (F₃))] and four CCC levels [(without CCC as control (C₀), application of 400 (C₁), 800 (C₂) and 1200 (C₃) mg/l)]. The results showed that water limitation decreased the chlorophyll-a, chlorophyll-b, total chlorophyll, carotenoid, stomata conductance, leaf area index (LAI) and relative water content of wheat, but activity of catalase (CAT), peroxidase (POD), polyphenol oxidase (PPO) enzymes and proline content were increased. Similar results were observed in CAT, POD and PPO activities due to bio fertilizers and CCC application. Besides the water limitation effects, CCC-treated plants displayed a significant decrease in stomata conductance and LAI. Generally, it was concluded that the application of bio fertilizers and CCC can be a proper tool for increasing wheat yield under water limitation.     

Salinity and drought interaction in wheat (Triticum aestivum L.) is affected by the genotype and plant growth stage

Salinity and drought are important agro-environmental problems occurring separately as well as together with the combined occurrence increasing with time due to climate change. Screening of bread wheat genotypes against salinity or drought alone is common; however, little information is available on the response of wheat genotypes to a combination of these stresses. This study investigates the response of a salt-resistant (SARC-1) and a salt-sensitive (7-Cerros) wheat genotype to drought at different growth stages under non-saline (ECe 2.1 dS m^?1) and saline soil (ECe 15 dS m^?1) conditions. Drought was applied by withholding water for 21 days at a particular growth stage viz. tillering, booting, and grain filling stages. At booting stage measurements regarding water relations, leaf ionic composition and photosynthetic attributes were made. At maturity grain yield and different yield, components were recorded. Salinity and drought significantly decreased grain yield and different yield components with a higher decrease in the case of combined stress of salinity × drought. The complete drought treatment (drought at tillering + booting + grain filling stages) was most harmful for wheat followed by drought at booting stage and grain filling–tillering stages, respectively. The salt-resistant wheat genotype SARC-1 performed better than the salt-sensitive genotype 7-Cerros in different stress treatments. A decrease in the water and turgor potentials, photosynthetic and transpiration rates, stomatal conductance, leaf K⁺, and increased leaf Na⁺ were the apparent causes of growth and yield reduction of bread wheat due to salinity, drought, and salinity × drought.     

Effects of applying stem-shortening plant growth regulators to leaves on root elongation by seedlings of wheat, oat and barley: mediation by ethylene

Several plant growth regulators (PGRs) commonly used in practicalfarming to restrict shoot height and control lodging were examined for theirimpact on root growth in naturally short or tall cultivars of barley (cvs.Kymppi and Saana), oat (cvs. Veli and Pal), and wheat (cvs. Mahti and Tjalve).The possible involvement of ethylene in the responses was also examined. Shootswere sprayed at the two-leaf stage with the gibberellin biosynthesis inhibitorsCycocel (chlormequat chloride) (CCC) or Moddus (Trinexapac-ethyl) (TE), or withthe ethylene-releasing agent Cerone (ethephon) (ETH) at 0, 0.1, 1, 10 or 50times the recommended agricultural rate (RR). Root elongation and ethyleneproduction by roots or shoots were unaffected by CCC at all application ratesorby TE at ×0.1 or ×1.0 RR. At ×10 and ×50 RR, TE wasinhibitory to root extension but did not increase ethylene biosynthesis bytheseroots or the shoots. ETH at ×0.1 or ×1.0 RR did not affect rootextension or ethylene production in roots or shoots. At all higher rates ofapplication ETH stimulated ethylene production strongly in shoots and roots ofall three species, while root elongation was retarded severely in barley,moderately in oat and only slightly in wheat. These differences in elongationresponse are attributed to differences in sensitivity to ethylene released byethephon. Accordingly, root elongation in wheat was only slightly affected whenethylene gas was supplied at concentrations up to 100 ppm for 3d. In contrast, root elongation in barley was strongly inhibitedbyethylene, with oat demonstrating an intermediate responsiveness.     

Effects of mycorrhizal infection on drought tolerance and recovery in safflower and wheat

The influence of arbuscular mycorrhizal fungi on drought tolerance and recovery was studied in safflower (Carthamus tinctorius L.) and wheat (Triticum aestivum L.). Plants were grown with and without the mycorrhizal fungus, Glomus etunicatum Becker & Gerd., in nutrient-amended soil under environmentally-controlled conditions to yield mycorrhizal and nonmycorrhizal with similar leaf areas, root length densities, dry weights, and adequate tissue phosphorus. When drought stress was induced, mycorrhizal infection did not affect changes in leaf water, osmotic or pressure potentials, or osmotic potentials of leaf tissue rehydrated to full turgor in either safflower or wheat. Furthermore, in safflower, infection had little effect on drought tolerance as indicated by the level of leaf necrosis. Mycorrhizal wheat plants, however, had less necrotic leaf tissue than uninfected plants at moderate levels of drought stress (but not at severe levels) probably due to enhanced phosphorus nutrition. To determine the effects of infection on drought recovery, plants were rewatered at a range of soil water potentials from –1 to –4 MPa. We found that although safflower tended to recover more slowly from drought after rewatering than wheat, mycorrhizal infection did not directly affect drought recovery in either plant species. Daily water use after rewatering was reduced and was correlated to the extent that leaves were damaged by drought stress in both plant species, but was not directly influenced by the mycorrhizal status of the plants.     

Effects of Mineral Nutrition on Water Status and Osmotic Adjustment in Spring wheat under Different Moisture Conditions

The Ψw and RWC of the leaves of spring wheat at low level of nutrition were higher than those of the spring wheat leaves at high level during the slow drought period. Therefore, the plants at high nutrition level were more sensitive to drought. High mineral nutrition did not enhance the maximum osmotic adjustment but changed the regulatory process of osmotic adjustment under drought condition. During moderate to nearly severe drought period higher osmotic adjustment was shown in the leaves of spring wheat at high nutrition level. The major solute contributed to osmotic adjustment in the leaves of the spring wheat was soluble sugar, mostly reducing sugar.     

Effect of CCC and glycine betaine on growth and growth-substance content of primary leaves of dwarf Frenchbean (Phaseolus vulgaris L.)

CCC (2-chloroethyltrimethylammonium chloride) decreased growth of primary leaves and stem internodes above the hypocotyl of dwarf French bean, probably because it decreased the amount of auxin produced by interaction between tryptophan and phenolic esters in the primary leaves. Growth of leaves was less affected than that of stems, and the gibberellin content of the primary leaves, previously shown to be associated with their expansion, was unaffected by CCC. CCC delayed death of the primary leaves, the breakdown of chlorophyll, and the increase in auxin associated with death of bean leaves. CCC had less effect on the growth of leaf discs, probably because they are not sites of growth-substance production. Glycine betaine, a quaternary ammonium compound similar to CCC, did not affect growth of bean plants, their metabolism of growth substances or the longevity of their primary leaves, but inhibited growth of leaf discs, probably directly.     

Biological changes in the rhizosphere of wheat after foliar application of chlorocholinechloride,urea and 4-chloro-2-methylphenoxyacetic acid

In field experiments wheat in the phase of shooting was sprayed with solutions of chlorocholinechloride (CCC) and urea, CCC and ammonium salt MCPA (Aminex) or CCC, urea and Aminex. The effect of the treatment on dry weight of overground parts of wheat, number of bacteria, production of carbon dioxide, urease activity and content of ammonium in the rhizosphere soil was investigated. In all cases evolution of carbon dioxide in the rhizosphere soil was higher than that in the control soil. Highest numbers of bacteria were found in the rhizosphere soil of plants treated with urea, the herbicide and their mixtures. Content of ammonium was higher in the control soil than in the rhizosphere soils, the urease activity was highest in the rhizosphere soil of plants treated with the solution of the herbicide and with the combination of the herbicide with urea.     

Endogenous auxins in apical buds ofChenopodium rubrum L. after application of (2-chloroethyl) trimethylammonium chloride (CCC)

Apical buds ofChenopodium rubrum from plants treated with CCC contain more endogenous auxins than buds from control plants, the level of these compounds increasing with the application of rising concentrations of the retardant. An especially marked increase was observed in the level of substance “X” which on chromatographic separation runs in the zone of tryptamine or its derivative. Since it has been shown in previous experiments that the inhibitory effect of CCC on flowering ofChenopodium rubrum may be reversed by indole-3-acetic acid (IAA) it is believed that the increase in auxins after application of CCC does not concern biologically active substances immediately available to the plant. It seems more likely that inactive precursors are involved which cannot be converted to the active substance in the presence of CCC, possibly due to blocking of the pertinent enzyme. If we assume that the wheat coleoptile used in the auxin bioassay in our experiments contains the pertinent enzyme it might convert the inactive precursors to active substances and, therefore, exhibit a growth stimulation even though the substances concerned would not necessarily be active in the buds from which they were extracted.