Gene expression changes in response to drought stress in <Emphasis Type="Italic">Citrullus colocynthis</Emphasis>

Citrullus colocynthis (L.) Schrad, closely related to watermelon, is a member of the Cucurbitaceae family. This plant is a drought-tolerant species with a deep root system, widely distributed in the Sahara-Arabian deserts in Africa and the Mediterranean region. cDNA amplified fragment length polymorphism (cDNA-AFLP) was used to study differential gene expression in roots of seedlings in response to a 20% polyethylene glycol-(PEG8000) induced drought stress treatment. Eighteen genes which show similarity to known function genes were confirmed by quantitative relative (RQ) real-time RT-PCR to be differentially regulated. These genes are involved in various abiotic and biotic stress and developmental responses. Dynamic changes with tissue-specific pattern were detected between 0 and 48 h of PEG treatment. In general, the highest induction levels in roots occurred earlier than in shoots, because the highest expression was detected in roots following 4 and 12 h, in shoots following 12 and 48 h of drought. These drought-responsive genes were also affected by the plant hormones abscisic acid (ABA), salicylic acid (SA), or jasmonic acid (JA), indicating an extensive cross-talk between drought and plant hormones. Collectively, these results will be useful to explore the functions of these multiple signal-inducible genes for unveiling the relationship and crosstalk between different signaling pathways.     

Drought-induced changes in photosynthetic membranes of two wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) cultivars

Two wheat (Triticum aestivum L.) cultivars contrasting in architectonics and differing in drought resistance, Azamatli-95 (short stature, vertically oriented small leaves, drought-tolerant) and Giymatli-2/17 (short stature, broad and drooping leaves, drought-sensitive), were studied. It was found that the content of CP I (115 kDa) and 63-kDa apoprotein P700 and also LHC II polypeptides increases slightly in the drought-resistant cv. Azamatli-95 under extreme water supply limitation, while their content decreases in drought-sensitive cv. Giymatli-2/17. The intensity of synthesis of α- and β-subunits of CF₁ (55 and 53.5 kDa) and 33–30.5 kDa proteins also decreases in the sensitive genotype. The intensity of short wavelength peaks at 687 and 695 nm sharply increases in the fluorescence spectra (77K) of chloroplasts from Giymatli-2/17 under water deficiency, and there is a stimulation of the ratio of fluorescence band intensity F687/F740. After exposure to drought, cv. Giymatli-2/17 shows a larger reduction in the actual PS II photochemical efficiency of chloroplasts than cv. Azamatli-95. Published in Russian in Biokhimiya, 2009, Vol. 74, No. 8, pp. 1109–1116.     

Molecular cloning and characterization of a novel glyoxalase I gene <Emphasis Type="Italic">TaGly I</Emphasis> in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Methylglyoxal is a kind of poisonous metabolite that can react with RNA, DNA and protein, which generally results in a number of side advert effects to cell. Glyoxalase I is a member of glyoxalase system that can detoxify methylglyoxal. An EST encoding a glyoxalase I was isolated from a SSH (suppression subtractive hybridization)-cDNA library of wheat spike inoculated by Fusarium graminearum. The corresponding full length gene, named TaGly I, was cloned, sequenced and characterized. Its genomic sequence consists of 2,719 bp, including seven exons and six introns, and its coding sequence is 929 bp with an open reading frame encoding 291 amino acids. Sequence alignment showed that there were two glyoxalase I domains in the deduced protein sequence. By using specific primers, TaGly I was mapped to chromosome 7D of wheat via a set of durum wheat ‘Langdon’ D-genome disomic-substitution lines. The result of Real-time quantitative polymerase chain reaction demonstrated that TaGly I was induced by the inoculation of Fusarium graminearum in wheat spikes. Additionally, it was also induced by high concentration of NaCl and ZnCl₂. When TaGly I was overexpressed in tobacco leaves via Agrobacterium tumefaciens infection, the transgenic tobacco showed stronger tolerance to ZnCl₂ stress relative to transgenic control with GFP. The above facts indicated that TaGly I might play a role in response to diverse stresses in plants.     

The endophytic fungi from wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

In order to study the species composition of endophytes from wheat healthy plants in Buenos Aires Province (Argentina) and to determine their infection frequencies from leaves, stems, glumes and grains, wheat plants were collected from five cultivars at five growth stages from crop emergence to harvest. A total of 1,750 plant segments (leaves, stems, glumes and grains) were processed from the five wheat cultivars at five growth stages, and 722 isolates of endophytic fungi recovered were identified as 30 fungal genera. Alternaria alternata, Cladosporium herbarum, Epicoccum nigrum, Cryptococcus sp., Rhodotorula rubra, Penicillium sp. and Fusarium graminearum were the fungi that showed the highest colonization frequency (CF%) in all the tissues and organs analysed. The number of taxa isolated was greater in the leaves than those in the other organs analysed.     

The physiological characteristics of the low canopy temperature wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) genotypes under simulated drought condition

Crop canopy temperature reflects the interactions among plants, soil and atmosphere. Through a long-term investigation into physiological characteristics of different canopy temperature wheat, it is found that some wheat exist a high canopy temperature while others a low one, which was closely correlated with their corresponding performance. Under simulated drought conditions, the physiological aspects of different canopy temperature wheat, such as leaf functional duration, chlorophyll content, activities of superoxide dismutase, protein content, transpiration rate, net photosynthesis rate, etc., were investigated, the result showed that wheat with low canopy temperature could maintain superiority to wheat with high canopy temperature in those physiological traits. Therefore, the low canopy temperature in wheat could be used as an index to evaluate physiological capacities of wheat under drought conditions and also as a useful marker for wheat breeding for drought tolerance.     

Effects of colchicine on anther and microspore culture of bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

The aim of this work was to study the effects of colchicine application on chromosome doubling and androgenic response in anther and microspore culture of different bread wheat genotypes. Colchicine was applied during a mannitol stress pretreatment or during the first 48 h of culture at concentrations of 0, 150 and 300 mg l⁻¹. When colchicine was applied during stress pretreatment, the percentage of doubling depended on genotype and concentration. A significant increase in doubling was observed with 300 mg l⁻¹ in the low androgenic responding cv. Caramba. Colchicine incorporation during the first hours of culture improved percentage of doubling in all genotypes, in both anther and microspore culture. Application of 300 mg l⁻¹ colchicine improved the percentage of doubling in the two low responding genotypes, to 118% of control in DH24033, and 75% in Caramba in microspore and anther culture, respectively. Concerning the androgenic response, the effect of colchicine on embryo formation and percentage of green plants depended on the genotype and on the culture method. In cv. Pavon, a 2- and a 3-fold increase in percentage of embryogenesis and green plants, respectively, were obtained with 300 mg l⁻¹ colchicine in microspore culture. However, no significant differences in these two variables were observed in anther culture. The number of green doubled haploid (DH) plants reflects the index of success of the procedure. Regardless of the culture method, when colchicine was incorporated during the first hours of culture, the number of green DH plants increased significantly in three of four genotypes. These results confirm the usefulness of colchicine application during the first hours of culture in wheat breeding programs.     

Antioxidative protection and proteolytic activity in tolerant and sensitive wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) varieties subjected to long-term field drought

Field drought studies were performed in order to assess oxidative stress, proteolytic activity and yield loss under natural stress conditions. Flag leaves of two drought-tolerant (Yantar and Zlatitsa) and two drought-sensitive (Miziya and Dobrudjanka) winter wheat varieties were analyzed. Stress intensity was assessed by relative electrolyte leakage and proline accumulation. Senescence progression was followed by loss of chlorophyll and protein. Lipid peroxidation, H₂O₂ content, activities of superoxide dismutase (SOD), catalase (CAT), and non-specific peroxidase (GPX) isoforms, as well as proteolytic activities were analyzed from heading throughout grain filling. Weakening of membrane integrity and oxidative damage to lipids were more pronounced in the sensitive varieties under field drought. The activities of Fe- and Cu/Zn SOD isoforms decreased in the controls, but remained high in drought-treated plants. The activities of MnSOD isoforms and CAT were enhanced towards grain filling, especially in the sensitive varieties under drought. GPX activities were rised under drought but progressively diminished. Accelerated senescence under field drought was linked to higher proteolytic activity with variety specific differences in the protease response, but without a clear correlation to drought resistance or sensitivity. Field drought led to higher oxidative stress more pronounced for drought sensitive varieties, especially during the grain filling period.     

Genotypic variation in drought stress response and subsequent recovery of wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Three wheat (Triticum aestivum L.) genotypes, Sadovo, Katya and Prelom, with different tolerance to drought were comparatively evaluated in terms of leaf respiratory responses to progressing dehydration and consecutive rewatering. Under drought stress, the respiration of all varieties gradually decreased, as the drought-tolerant Katya showed the most pronounced decline at earlier stages of dehydration. When water stress intensified, this genotype gave relatively stable respiration rates compared with the drought-sensitive varieties. Additionally, dehydrated Katya leaves displayed lower stomatal conductance and higher photosynthesis values, which resulted in greater water use efficiency during the dehydration period. Combination of drought stress and short-term changes in leaf temperature also induced genotype-specific response that differed from the response to drought only. Over the whole temperature range, the leaves of Katya exposed to dehydration for 14 days, showed higher respiration rates compared to the drought-sensitive varieties. The sensitive varieties maintained higher respiration rates under control conditions and mild dehydration, and very low rates under severe drought. In Katya, respiration and photosynthesis were fully restored from the stress within the first day of rewatering. The drought-sensitive genotypes displayed a considerably slower recovering capacity. The results are discussed in terms of possible physiological mechanisms underlying plant tolerance to drought.     

The parameters of grain filling and yield components in common wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) and durum wheat (<Emphasis Type="Italic">Triticum turgidum</Emphasis> L. var. <Emphasis Type="Italic">durum</Emphasis>)

Final grain dry weight, a component of yield in wheat, is dependent on the duration and the rate of grain filling. The purpose of the study was to compare the grain filling patterns between common wheat, (Triticum aestivum L.), and durum wheat, (Triticum turgidum L. var. durum), and investigate relationships among grain filling parameters, yield components and the yield itself. The most important variables in differentiating among grain filling curves were final grain dry weight (W) for common wheat genotypes and grain filling rate (R) for durum wheat genotypes; however, in all cases the sets of variables important in differentiating among grain filling curves were extended to either two or all three parameters. Furthermore, in one out of three environmental conditions and for both groups of genotypes, the most important parameter in the set was grain filling duration (T). It indicates significant impact of environmental conditions on dry matter accumulation and the mutual effect of grain filling duration and its rate on the final grain dry weight. The medium early anthesis date could be associated with further grain weight and yield improvements in wheat. Grain filling of earlier genotypes occurs in more temperate environments, which provides enough time for gradual grain fill and avoids the extremes of temperature and the stress of dry conditions.     

Freezing of dehydrated calli of spring wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) in liquid nitrogen and their morphogenetic potential

A cryopreservation method developed earlier was modified for freezing of calli derived from mature embryos of four spring wheat (Triticum aestivum L.) pure lines. The effects of particular stages of cryopreservation protocol on water content, number of calli recovering growth, and rate of morphogenesis were analyzed. Regrowth was observed in 90.5–100% of calli after dehydration and in 93.3–100% after freezing-thawing. Dehydration, but not freeze-thawing significantly decreased the frequency of morphogenetic variation.     

Overexpression of <Emphasis Type="Italic">AtHDG11</Emphasis> enhanced drought tolerance in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Drought is one of the major abiotic stresses restricting the yield of wheat (Triticum aestivum L.). Breeding wheat varieties with drought tolerance is an effective and durable way to fight against drought. Here we reported introduction of AtHDG11 into wheat via Agrobacterium-mediated transformation and analyzed the morphological and physiological characteristics of T2 generation transgenic lines under drought stress. With drought treatment for 30 days, transgenic plants showed significantly improved drought tolerance. Compared with controls, the transgenic lines displayed lower stomatal density, lower water loss rate, more proline accumulation and increased activities of catalase and superoxide dismutase. Without irrigation after booting stage, the photosynthetic parameters, such as net photosynthesis rate, water use efficiency and efficiency of excitation energy, were increased in transgenic lines, while transpiration rate was decreased. Moreover, the kernel yield of transgenic lines was also improved under drought condition. Taken together, our data demonstrate that AtHDG11 has great potential in genetic improvement of drought tolerance of wheat.     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of ten threatened species of <Emphasis Type="Italic">Mammillaria</Emphasis> (Cactaceae)

Mammillaria species are the most numerous within Cactaceae family, and some of them are threatened with extinction as a result of human activities. In this work, results of in vitro propagation are presented for ten Mammillaria species, testing 20 combinations of indole-3-acetic acid (IAA) and kinetin. Best results on shoot formation were obtained using kinetin at two levels: 27.9 and 46.5 μM. All IAA levels tested were able to induce de novo shoot formation in M. bocasana, M. densispina, M. hahniana, M. hutchisoniana, M. orcutii, M. pectinifera, M. perbella, M. picta, M. rhodantha, and M. zephyranthoides. Depending on the IAA level tested, four responding groups were observed concerning their highest shoot-formation number. For all species, the highest average of shoot formation was achieved with 5.7:46.5 or 11.4:46.5 μM IAA/kinetin, yielding 4.8 and 4.7 shoots per explant, respectively, in 60 d. Rooting of regenerated shoots was achieved by leaving the explants in their shoot-induction medium or transferring them to half-strength MS medium. Hardening of regenerated plants was successfully achieved by planting them in peat moss substrate after a desiccation treatment at room temperature for 3 d.     

Cloning and characterization of microRNAs from wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Background  

MicroRNAs (miRNAs) are a class of small, non-coding regulatory RNAs that regulate gene expression by guiding target mRNA cleavage or translational inhibition. So far, identification of miRNAs has been limited to a few model plant species, such as Arabidopsis, rice and Populus, whose genomes have been sequenced. Wheat is one of the most important cereal crops worldwide. To date, only a few conserved miRNAs have been predicted in wheat and the computational identification of wheat miRNAs requires the genome sequence, which is unknown.