Mepiquat chloride seed treatment and germination temperature effects on cotton growth, nutrient partitioning, and water use efficiency

Cotton seed (Gossypium hirsutum L. cv. Stoneville 825), treated with 0, 0.2, 1.0, and 2.0 g active ingredient (a.i.) mepiquat chloride (MC) kg^–1, was evaluated for the effect of MC on early plant growth. Emergence rate and total emergence of MC-treated seed and control were similar regardless of germination temperature. However, the number of leaves and squares and the dry weight of leaves, stems, and roots for hydroponically grown cotton plants were significantly lower at lower germination temperatures (15°C for 3 day/30°C for 1 day and 15°C for 4 days) than at higher germination temperatures (30°C for 4 days and 30°C for 3 days/15°C for 1 day). All MC treatments significantly decreased the number of nodes, leaves, and squares, as well as dry weight of leaves, stems, and roots, as compared to control plants at 28 days after emergence. MC seed treatments also significantly reduced plant height and total leaf area compared to controls. Water-use efficiency (WUE) was significantly lower for the 1.0 g a.i. MC treatment than for control plants. In general, the highest rate of MC seed treatment resulted in greater concentrations of calcium, phosphorus, and nitrogen in plant leaves and stems and also in greater concentrations of magnesium, phosphorus, and nitrogen in roots than in controls.     

The cyclic nucleotide gated cation channel AtCNGC10 traffics from the ER via Golgi vesicles to the plasma membrane of Arabidopsis root and leaf cells

Background  

The cyclic nucleotide-gated ion channels (CNGCs) maintain cation homeostasis essential for a wide range of physiological processes in plant cells. However, the precise subcellular locations and trafficking of these membrane proteins are poorly understood. This is further complicated by a general deficiency of information about targeting pathways of membrane proteins in plants. To investigate CNGC trafficking and localization, we have measured Atcngc5 and Atcngc10 expression in roots and leaves, analyzed AtCNGC10-GFP fusions transiently expressed in protoplasts, and conducted immunofluorescence labeling of protoplasts and immunoelectron microscopic analysis of high pressure frozen leaves and roots.     

The expression and anaerobic induction of alcohol dehydrogenase in cotton

The alcohol dehydrogenase (ADH) system in cotton is characterized, with an emphasis on the cultivated allotetraploid speciesGossypium hirsutum cv. Siokra. A high level of ADH activity is present in seed of Siokra but quickly declines during germination. When exposed to anaerobic stress the level of ADH activity can be induced several fold in both roots and shoots of seedlings. Unlike maize andArabidopsis, ADH activity can be anaerobically induced in mature green leaves. Three major ADH isozymes were resolved in Siokra, and it is proposed that two genes,Adh1 andAdh2, are coding for these three isozymes. The genes are differentially expressed. ADH1 is predominant in seed and aerobically grown roots, while ADH2 is prominent in roots only after anaerobic stress. Biochemical analysis demonstrated that the ADH enzyme has a native molecular weight of approximately 81 kD and a subunit molecular weight of approximately 42 kD, thus establishing that ADH in cotton is able to form and is active as dimers. Comparisons of ADH activity levels and isozyme patterns between Siokra and other allotetraploid cottons showed that the ADH system is highly conserved among these varieties. In contrast, the diploid species of cotton all had unique isozyme patterns.This work was generously supported by an Australian Cotton Research Council Postgraduate Studentship.     

Mepiquat chloride seed treatment and germination temperature effects on cotton growth,nutrient partitioning,and water use efficiency

Cotton seed (Gossypium hirsutum L. cv. “Stoneville 825”), treated with 0, 0.2, 1.0, and 2.0 g active ingredient (a.i.) mepiquat chloride (MC) kg^?1, was evaluated for the effect of MC on early plant growth. Emergence rate and total emergence of MC-treated seed and control were similar regardless of germination temperature. However, the number of leaves and squares and the dry weight of leaves, stems, and roots for hydroponically grown cotton plants were significantly lower at lower germination temperatures (15°C for 3 day/30°C for 1 day and 15°C for 4 days) than at higher germination temperatures (30°C for 4 days and 30°C for 3 days/15°C for 1 day). All MC treatments significantly decreased the number of nodes, leaves, and squares, as well as dry weight of leaves, stems, and roots, as compared to control plants at 28 days after emergence. MC seed treatments also significantly reduced plant height and total leaf area compared to controls. Water-use efficiency (WUE) was significantly lower for the 1.0 g a.i. MC treatment than for control plants. In general, the highest rate of MC seed treatment resulted in greater concentrations of calcium, phosphorus, and nitrogen in plant leaves and stems and also in greater concentrations of magnesium, phosphorus, and nitrogen in roots than in controls.     

Survey of indigenous bacterial endophytes from cotton and sweet corn

The genotypic diversity of indigenous bacterial endophytes within stems and roots of sweet corn (Zea mays L.) and cotton (Gossypium hirsutum L.) was determined in field trials throughout one growing season. Strains were isolated from surface-disinfested tissues and identified by fatty acid analysis. Gram-negative bacteria comprised 70.5% of the endophytic bacteria and 27 of the 36 genera identified. The most frequently isolated groups from sweet corn roots, were Burkholderia pickettii and Enterobacter spp.; from sweet corn stems, Bacillus megaterium. Bacterial genera present in sweet corn roots were also generally present in sweet corn stems. However, Burkholderia gladioli, Burkholderia solanacearum and Enterobacter cloacae were isolated much more frequently from sweet corn roots than stems, whereas Methylobacterium spp. were found more frequently in sweet corn stems than roots. Agrobacterium radiobacter, Serratia spp. and Burkholderia solanacearum, were the most frequently isolated groups from cotton roots; and Bacillus megaterium and Bacillus pumilus from cotton stems. Acinetobacter baumannii and Arthrobacter spp. were present in cotton stems but not in cotton roots. There were 14 taxonomic groups present in cotton roots that were not in cotton stems; all but one were Gram-negative. These included, Agrobacterium radiobacter, Bacillus megaterium, Bacillus pumilus, Enterobacter asburiae, Pseudomonas chlororaphis, Serratia spp. and Staphylococcus spp. Rhizobium japonicum and Variovorax paradoxus were isolated, almost exclusively, from the roots of both crops. Bacterial taxa present in both sweet corn and cotton early in the season were generally present late in the season. The diversity of bacteria was greater in roots than stems for each crop.     

Identification of the family of aquaporin genes and their expression in upland cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis>L.)

Background  

Cotton (Gossypium spp.) is produced in over 30 countries and represents the most important natural fiber in the world. One of the primary factors affecting both the quantity and quality of cotton production is water. A major facilitator of water movement through cell membranes of cotton and other plants are the aquaporin proteins. Aquaporin proteins are present as diverse forms in plants, where they function as transport systems for water and other small molecules. The plant aquaporins belong to the large major intrinsic protein (MIP) family. In higher plants, they consist of five subfamilies including plasma membrane intrinsic proteins (PIP), tonoplast intrinsic proteins (TIP), NOD26-like intrinsic proteins (NIP), small basic intrinsic proteins (SIP), and the recently discovered X intrinsic proteins (XIP). Although a great deal is known about aquaporins in plants, very little is known in cotton.     

Phosphoenol-pyruvate-carboxylase activity in cotton and Sorghum seeds and its relation to seedling development

Cotton (Gossypium hirsutum) seeds and Sorghum vulgare caryopses are able to incorporate CO₂ through a PEP-carboxylating enzyme (EC 4.1.1.38). The enzyme activity is optimal at pH 8.2 and is unaffected by ATP, GDP or acetyl CoA. The partially purified cotton enzyme is stimulated by inorganic phosphate with an apparent Km of 0.3 mM. The enzymes from both cultivars are inhibited by pyrophosphate, malate, and aspartate but not by succinate. Kinetic studies for Sorghum and cotton seed enzymes show apparent Km values for carbonate of 5 mM and 1.2 mM and for PEP of 36 M and 5 mM, respectively. The V_max values are 90 and 3.3 nmol min^-1 mg protein^-1, respectively.A two-fold increase in the enzyme activity from cotton seeds occurs after 2 h under laboratory germination conditions after which the activity drops sharply to 1/3 of the original activity after 5 h imbibition. No such change was observed in Sorghum caryopses enzyme. A correlation between PEP-carboxylase activity and seed vigor in both cultivars was demonstrated.Abbreviations GOT glutamicoxaloacetic-transaminase - MDH malic dehydrogenase-NADH₂ - RH relative humidity     

Blue and UV LED-induced fluorescence in cotton foreign matter

Background

Cotton is an important domesticated fiber used to manufacture a variety of products and industrial goods. During harvesting with cotton strippers and cotton pickers, it is contaminated with foreign matter from botanical and non-botanical sources which adversely affect the quality and consistency of cotton, and therefore reduces its market value. To improve the current grading done by the High Volume Instrument (HVI) and human inspectors, it was explored whether fluorescence imaging can be used for cotton foreign matter detection.

Results

Eight types of botanical foreign matter (bark, bract, brown leaf, green leaf, hull, seed coat, seed, stem), and four types of non-botanical foreign matter (paper, twine, plastic bale packaging, plastic bag) were subjected to a fluorescence spectroscopy analysis to determine their optimal excitation and emission wavelength range. Matrix 3D scans were performed in the excitation range from 300 nm to 500 nm, and emission range from 320 nm to 700 nm, and the results indicated the photo-excitable fluorescence in the aforementioned excitation range for all the selected foreign matter categories. Consequently, the blue and the UV LEDs were selected as the excitation sources. The blue LED light provided optimal excitation light for bark, brown leaf, bract, green leaf, hull, and stem, while the UV LED light provided optimal excitation light for paper, plastic bag, plastic packaging, seed, seed coat, and twine.

Conclusions

UV and blue light induces fluorescence in 12 types of botanical and non-botanical cotton foreign matter. An imaging apparatus with blue and UV light excitation sources, and a consumer grade SLR camera was successfully developed to capture and characterize fluorescent images of cotton foreign matter. Based on the results, fluorescent imaging could be a promising method for cotton foreign matter detection. Future studies will focus on the classification of cotton foreign matter categories and to further refine the image processing sequence.

     

Proteome analysis of Norway maple (Acer platanoides L.) seeds dormancy breaking and germination: influence of abscisic and gibberellic acids

Background  

Seed dormancy is controlled by the physiological or structural properties of a seed and the external conditions. It is induced as part of the genetic program of seed development and maturation. Seeds with deep physiological embryo dormancy can be stimulated to germinate by a variety of treatments including cold stratification. Hormonal imbalance between germination inhibitors (e.g. abscisic acid) and growth promoters (e.g. gibberellins) is the main cause of seed dormancy breaking. Differences in the status of hormones would affect expression of genes required for germination. Proteomics offers the opportunity to examine simultaneous changes and to classify temporal patterns of protein accumulation occurring during seed dormancy breaking and germination. Analysis of the functions of the identified proteins and the related metabolic pathways, in conjunction with the plant hormones implicated in seed dormancy breaking, would expand our knowledge about this process.     

Multiplex real‐time PCR assays for detection of four seedborne spinach pathogens

Aims

To develop multiplex TaqMan real‐time PCR assays for detection of spinach seedborne pathogens that cause economically important diseases on spinach.

Methods and Results

Primers and probes were designed from conserved sequences of the internal transcribed spacer (for Peronospora farinosa f. sp. spinaciae and Stemphylium botryosum), the intergenic spacer (for Verticillium dahliae) and the elongation factor 1 alpha (for Cladosporium variabile) regions of DNA. The TaqMan assays were tested on DNA extracted from numerous isolates of the four target pathogens, as well as a wide range of nontarget, related fungi or oomycetes and numerous saprophytes commonly found on spinach seed. Multiplex real‐time PCR assays were evaluated by detecting two or three target pathogens simultaneously. Singular and multiplex real‐time PCR assays were also applied to DNA extracted from bulked seed and single spinach seed.

Conclusions

The real‐time PCR assays were species‐specific and sensitive. Singular or multiplex real‐time PCR assays could detect target pathogens from both bulked seed samples as well as single spinach seed.

Significance and Impact of the Study

The freeze‐blotter assay that is currently routinely used in the spinach seed industry to detect and quantify three fungal seedborne pathogens of spinach (C. variabile, S. botryosum and V. dahliae) is quite laborious and takes several weeks to process. The real‐time PCR assays developed in this study are more sensitive and can be completed in a single day. As the assays can be applied easily for routine seed inspections, these tools could be very useful to the spinach seed industry.     

Analysis of gene expression in cotton fiber initials

Background  

Cotton (Gossypium hirsutum L.) fibers are trichomes that initiate from the ovule epidermis. Little is known about the developmental pathway causing fiber to differentiate from ovular epidermal cells even though limits on the number of cells that differentiate into fiber will limit yield.     

Investigating seed dormancy in cotton (Gossypium hirsutum L.): understanding the physiological changes in embryo during after‐ripening and germination

• The dormancy of seeds of upland cotton can be broken during dry after‐ripening, but the mechanism of its dormancy release remains unclear.
• Freshly harvested cotton seeds were subjected to after‐ripening for 180 days. Cotton seeds from different days of after‐ripening (DAR) were sampled for dynamic physiological determination and germination tests. The intact seeds and isolated embryos were germinated to assess effects of the seed coat on embryo germination. Content of H₂O₂ and phytohormones and activities of antioxidant enzymes and glucose‐6‐phosphate dehydrogenase were measured during after‐ripening and germination.
• Germination of intact seeds increased from 7% upon harvest to 96% at 30 DAR, while embryo germination improved from an initial rate of 82% to 100% after 14 DAR. Based on T₅₀ (time when 50% of seeds germinate) and germination index, the intact seed and isolated embryo needed 30 and 21 DAR, respectively, to acquire relatively stable germination. The content of H₂O₂ increased during after‐ripening and continued to increase within the first few hours of imbibition, along with a decrease in abscisic acid (ABA) content. A noticeable increase was observed in gibberellic acid content during germination when ABA content decreased to a lower level. Coat removal treatment accelerated embryo absorption of water, which further improved the accumulation of H₂O₂ and changed peroxidase content during germination.
• For cotton seed, the alleviation of coat‐imposed dormancy required 30 days of after‐ripening, accompanied by rapid dormancy release (within 21 DAR) in naked embryos. H₂O₂ acted as a core link between the response to environmental changes and induction of other physiological changes for breaking seed dormancy.

     

A fungal endophyte defensive symbiosis affects plant-nematode interactions in cotton

Background and aims

Most investigations of fungi as nematode antagonists have focused on their interactions with nematodes in the soil. This study tested a foliar-isolated endophytic Phialemonium inflatum for its effects against the root-knot nematode as an endophyte in cotton using a seed treatment inoculation.

Methods

Cotton seeds were inoculated with P. inflatum spore suspensions prior to planting. Nematode infection and reproduction were quantified at Day 12 and 6 weeks after nematode egg inoculation, respectively. To establish whether the observed negative effects on nematodes were due to P. inflatum in the soil or as an endophyte in the plant, we also applied a soil fungicide treatment at the seedling stage to kill the fungi outside the plant.

Results

Persistent suppression of nematode penetration and galling, as well as subsequent reproduction, were observed in endophyte-treated plants independent of fungicide treatment, consistent with an endophytic mode of nematode suppression; and these negative effects did not depend on the concentration of fungal inoculum used to treat to the seed.

Conclusions

Our study highlights a novel role for P. inflatum as part of a plant-fungal defensive symbiosis in cotton, as well as the need for a broader understanding of endophyte-plant-nematode ecological interactions.

     

Genetic transformation of cotton with a harpin-encoding gene <Emphasis Type="Italic">hpa</Emphasis><Subscript><Emphasis Type="Italic">Xoo</Emphasis></Subscript> confers an enhanced defense response against different pathogens through a priming mechanism

Background  

The soil-borne fungal pathogen Verticillium dahliae Kleb causes Verticillium wilt in a wide range of crops including cotton (Gossypium hirsutum). To date, most upland cotton varieties are susceptible to V. dahliae and the breeding for cotton varieties with the resistance to Verticillium wilt has not been successful.