Live-cell imaging of the cytoskeleton in elongating cotton fibres.

Cotton (Gossypium hirsutum) fibres consist of single cells that grow in a highly polarized manner, assumed to be controlled by the cytoskeleton1-3. However, how the cytoskeletal organization and dynamics underpin fibre development remains unexplored. Moreover, it is unclear whether cotton fibres expand via tip growth or diffuse growth2-4. We generated stable transgenic cotton plants expressing fluorescent markers of the actin and microtubule cytoskeleton. Live-cell imaging revealed that elongating cotton fibres assemble a cortical filamentous actin network that extends along the cell axis to finally form actin strands with closed loops in the tapered fibre tip. Analyses of F-actin network properties indicate that cotton fibres have a unique actin organization that blends features of both diffuse and tip growth modes. Interestingly, typical actin organization and endosomal vesicle aggregation found in tip-growing cell apices were not observed in fibre tips. Instead, endomembrane compartments were evenly distributed along the elongating fibre cells and moved bi-directionally along the fibre shank to the fibre tip. Moreover, plus-end tracked microtubules transversely encircled elongating fibre shanks, reminiscent of diffusely growing cells. Collectively, our findings indicate that cotton fibres elongate via a unique tip-biased diffuse growth mode.

Genetics and evolution of MIXTA genes regulating cotton lint fiber development.

Cotton, with cellulose-enriched mature fibers, is the largest source of natural textiles. Through a map-based cloning strategy, we isolated an industrially important lint fiber development gene (Li3 ) that encodes an MYB-MIXTA-like transcription factor (MML) on chromosome D12 (GhMML4_D12). Virus-induced gene silencing or decreasing the expression of the GhMML4_D12 gene in n2 NSM plants resulted in a significant reduction in epidermal cell prominence and lint fiber production. GhMML4_D12 is arranged in tandem with GhMML3, another MIXTA gene responsible for fuzz fiber development. These two very closely related MIXTA genes direct fiber initiation production in two specialized cell forms: lint and fuzz fibers. They may control the same metabolic pathways in different cell types. The MIXTAs expanded in Malvaceae during their evolution and produced a Malvaceae-specific family that regulates epidermal cell differentiation, different from the gene family that regulates leaf hair trichome development. Cotton has developed a unique transcriptional regulatory network for fiber development. Characterization of target genes regulating fiber production has provided insights into the molecular mechanisms underlying cotton fiber development and has allowed the use of genetic engineering to increase lint yield by inducing more epidermal cells to develop into lint rather than fuzz fibers.

Suppressing a Putative Sterol Carrier Gene Reduces Plasmodesmal Permeability and Activates Sucrose Transporter Genes during Cotton Fiber Elongation.

Plasmodesmata (PDs) play vital roles in cell-to-cell communication and plant development. Emerging evidence suggests that sterols are involved in PD activity during cytokinesis. However, whether sterols contribute to PD gating between established cells remains unknown. Here, we isolated GhSCP2D, a putative sterol carrier protein gene from elongating cotton (Gossypium hirsutum) fibers. In contrast to wild-type fiber PDs, which opened at 5 to 10 d postanthesis (DPA) and closed only at 15 to 25 DPA, plants with suppressed GhSCP2D expression had reduced sterol contents and closed PDs at 5 through 25 DPA The GhSCP2D-suppressed fibers exhibited callose deposition at the PDs, likely due to reduced expression of GhPdBG3-2A/D, which encodes a PD-targeting ß-1,3-glucanase. Both GhPdBG3-2A/D expression and callose deposition were sensitive to a sterol biosynthesis inhibitor. Moreover, suppressing GhSCP2D upregulated a cohort of SUT and SWEET sucrose transporter genes in fiber cells. Collectively, our results indicate that (1) GhSCP2D is required for GhPdBG3-2A/D expression to degrade callose at the PD, thereby contributing to the establishment of the symplasmic pathway; and (2) blocking the symplasmic pathway by downregulating GhSCP2D activates or increases the expression of SUTs and SWEETs, leading to the switch from symplasmic to apoplasmic pathways.

Effects of silicon treatment and inoculation with Fusarium oxysporum f. sp. vasinfectum on cellular defences in root tissues of two cotton cultivars.

BACKGROUND AND AIMS: Silicon has been shown to enhance the resistance of plants to fungal and bacterial pathogens. Here, the effect of potassium silicate was assessed on two cotton (Gossypium hirsutum) cultivars subsequently inoculated with Fusarium oxysporum f. sp. vasinfectum (Fov). Sicot 189 is moderately resistant whilst Sicot F-1 is the second most resistant commercial cultivar presently available in Australia. METHODS: Transmission and light microscopy were used to compare cellular modifications in root cells after these different treatments. The accumulation of phenolic compounds and lignin was measured. KEY RESULTS: Cellular alterations including the deposition of electron-dense material, degradation of fungal hyphae and occlusion of endodermal cells were more rapidly induced and more intense in endodermal and vascular regions of Sicot F-1 plants supplied with potassium silicate followed by inoculation with Fov than in similarly treated Sicot 189 plants or in silicate-treated plants of either cultivar not inoculated with Fov. Significantly more phenolic compounds were present at 7 d post-infection (dpi) in root extracts of Sicot F-1 plants treated with potassium silicate followed by inoculation with Fov compared with plants from all other treatments. The lignin concentration at 3 dpi in root material from Sicot F-1 treated with potassium silicate and inoculated with Fov was significantly higher than that from water-treated and inoculated plants. CONCLUSIONS: This study demonstrates that silicon treatment can affect cellular defence responses in cotton roots subsequently inoculated with Fov, particularly in Sicot F-1, a cultivar with greater inherent resistance to this pathogen. This suggests that silicon may interact with or initiate defence pathways faster in this cultivar than in the less resistant cultivar.

Leaf-based physiological, metabolic, and ultrastructural changes in cultivated cotton cultivars under cadmium stress mediated by glutathione.

Cadmium (Cd) pollution is present in the world over especially in the industrialized parts of the world. To reduce Cd accumulation in various crops especially food crops, alleviating agents such as reduced glutathione (GSH) can be applied, which are capable either to exclude or to sequester Cd contamination. This study investigated the leaf-based spatial distribution of physiological, metabolic, and microstructural changes in two cotton cultivars (Coker 312 and TM-1) under GSH-mediated Cd stress using single levels of Cd (50 µM) and GSH (50 µM) both separately and in mix along with control. Results showed that GSH revived the morphology and physiology of both cotton cultivars alone or in mix with Cd. Cd uptake was enhanced in all segments of leaf and whole leaf upon the addition of GSH. GSH alleviated Cd-induced reduction in the photosynthetic pigment compositions and chlorophyll a fluorescence parameters. Mean data of biomarkers (2,3,5-triphenyltetrazolium (TTC), total soluble protein (TSP), malondialdehyde (MDA), hydrogen peroxide (H2O2)) revealed the adverse effects of Cd stress on leaf segments of both cultivars, which were revived by GSH. The oxidative metabolism induced by Cd stress was profoundly influenced by exogenous GSH application. The microstructural alterations were mainly confined to chloroplastic regions of leaves under Cd-stressed conditions, which were greatly revived upon the GSH addition. As a whole, Cd stress greatly affected TM-1 as compared to Coker 312. These results suggest a positive role of GSH in alleviating Cd-mediated changes in different leaf sections of cotton cultivars.

Small interfering RNAs from bidirectional transcripts of GhMML3_A12 regulate cotton fiber development.

Natural antisense transcripts (NATs) are commonly observed in eukaryotic genomes, but only a limited number of such genes have been identified as being involved in gene regulation in plants. In this research, we investigated the function of small RNA derived from a NAT in fiber cell development. Using a map-based cloning strategy for the first time in tetraploid cotton, we cloned a naked seed mutant gene (N1 ) encoding a MYBMIXTA-like transcription factor 3 (MML3)/GhMYB25-like in chromosome A12, GhMML3_A12, that is associated with fuzz fiber development. The extremely low expression of GhMML3_A12 in N1 is associated with NAT production, driven by its 3' antisense promoter, as indicated by the promoter-driven histochemical staining assay. In addition, small RNA deep sequencing analysis suggested that the bidirectional transcriptions of GhMML3_A12 form double-stranded RNAs and generate 21-22 nt small RNAs. Therefore, in a fiber-specific manner, small RNA derived from the GhMML3_A12 locus can mediate GhMML3_A12 mRNA self-cleavage and result in the production of naked seeds followed by lint fiber inhibition in N1 plants. The present research reports the first observation of gene-mediated NATs and siRNA directly controlling fiber development in cotton.

Alleviation of lead-induced physiological, metabolic, and ultramorphological changes in leaves of upland cotton through glutathione.

Plants face changes in leaves under lead (Pb) toxicity. Reduced glutathione (GSH) has several functions in plant metabolism, but its role in alleviating Pb toxicity in cotton leaves is still unknown. In the present study, cotton seedlings (28 days old) were exposed to 500 µM Pb and 50 µM GSH, both alone and in combination, for a period of 10 days, in the Hoagland solution under controlled growth conditions. Results revealed Pb-induced changes in cotton's leaf morphology, photosynthesis, and oxidative metabolism. However, exogenous application of GSH restored leaf growth. GSH triggered build up of chlorophyll a, chlorophyll b, and carotenoid contents and boosted fluorescence ratios (F v/F m and F v/F 0). Moreover, GSH reduced the malondialdehyde (MDA), hydrogen peroxide (H2O2), and Pb contents in cotton leaves. Results further revealed that total soluble protein contents were decreased under Pb toxicity; however, exogenously applied GSH improved these contents in cotton leaves. Activities of antioxidant enzymes (catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), glutathione reductase (GR), and ascorbate peroxidase (APX)) were also increased by GSH application under Pb toxicity. Microscopic analysis showed that excess Pb shattered thylakoid membranes in chloroplasts. However, GSH stabilized ultrastructure of Pb-stressed cotton leaves. These findings suggested that exogenously applied GSH lessened the adverse effects of Pb and improved cotton's tolerance to oxidative stress.

Genotypic differences in photosynthetic performance, antioxidant capacity, ultrastructure and nutrients in response to combined stress of salinity and Cd in cotton.

Combined stress of salinity and heavy metal is a serious problem for crop production; however, physiological mechanisms of tolerance to such condition remain elusive in cotton. Here, we used two cotton genotypes differing in salt tolerance, to understand their response to salinity (NaCl) and cadmium (Cd) either alone or in combination (Cd + Na) via hydroponics. Results showed that salinity and/or Cd drastically reduced plant growth, chlorophyll content and photosynthesis, with greater effect observed in Zhongmian 41 (sensitive) than Zhong 9806 (tolerant). Although salinity and/or Cd induced malondialdehyde (MDA) accumulation in Zhongmian 41 at 5 and 10 days after treatment, MDA content remained unchanged in Zhong 9806, implying that Zhongmian 41 but not Zhong 9806 faced oxidative stress following exposure to salinity and/or Cd. Differential responses of antioxidant enzymes such as superoxide dismutase, guaiacol peroxidase, catalase and ascorbate peroxidase to Cd, NaCl and Cd + Na indicate genotype- and time course- dependent variations. In both genotypes, Cd content was decreased while Na concentration was increased under combined stress compared with Cd alone. Importantly, NaCl addition in Cd-containing medium caused remarkable reduction in Cd concentration, with the extent of reduction being also dependent on genotypes. The salt-tolerant genotypes had lower Na concentration than sensitive ones. Furthermore, obvious changes in leaf and root ultrastructure was observed under Cd, Na and Cd + Na stress, however Zhong 9806 was less affected compared with Zhongmian 41. These results may provide novel insight into the physiological mechanisms of Cd + Na stress tolerance in various cotton genotypes.

Action modes of recombinant endocellulase, EGA, and its domains on cotton fabrics.

OBJECTIVES: The action modes of an endocellulase, EGA, and its domains (CD9 and CBM3) during enzymatic treatment of cotton fabrics were investigated. RESULTS: EGA, CD9 and CBM3 had the binding capacity to cellulose substrates, of which the filter paper was the substrate with the strongest binding capacity. Analyses of scanning electronic microscopy indicated that EGA and its catalytic domain CD9 etched the surface of cotton fabrics and broke the fibers of long chains. On the other hand, the binding domain CBM3 only resulted in swelling of cotton fibers. Both EGA and its catalytic domain CD9 had minimal effect on the weight loss of cotton fabrics, whereas the effect of EGA and CD9 on the degree of polymerization and breaking strength was significant. After 12 h enzymatic action, the values of weight loss ratio for EGA and CD9 were 2.07 and 2.21 %, respectively, meanwhile the reductions in fabric strength were 27.04 % for EGA and 17.23 % for CD9. CONCLUSIONS: In contrast to the action of EGA and CD9, CBM3 showed no significant changes in terms of the weight loss ratio, degree of polymerization, and fabric strength.

Comparative proteomic and biochemical analyses reveal different molecular events occurring in the process of fiber initiation between wild-type allotetraploid cotton and its fuzzless-lintless mutant.

To explore lint fiber initiation-related proteins in allotetraploid cotton (Gossypium hirsutum L.), a comparative proteomic analysis was performed between wild-type cotton (Xu-142) and its fuzzless-lintless mutant (Xu-142-fl) at five developmental time points for lint fiber initiation from -3 to +3 days post-anthesis (dpa). Using two-dimensional gel electrophoresis (2-DE) combined with mass spectrometry (MS) analyses, 91 differentially accumulated protein (DAP) species that are related to fiber initiation were successfully identified, of which 58 preferentially accumulated in the wild-type and 33 species in the fl mutant. These DAPs are involved in various cellular and metabolic processes, mainly including important energy/carbohydrate metabolism, redox homeostasis, amino acid and fatty acid biosynthesis, protein quality control, cytoskeleton dynamics, and anthocyanidin metabolism. Further physiological and biochemical experiments revealed dynamic changes in the carbohydrate flux and H2O2 levels in the cotton fiber initiation process. Compared with those in the fl mutant, the contents of glucose and fructose in wild-type ovules sharply increased after anthesis with a relatively higher rate of amino acid biosynthesis. The relative sugar starvation and lower rate of amino acid biosynthesis in the fl mutant ovules may impede the carbohydrate/energy supply and cell wall synthesis, which is consistent with the proteomic results. However, the H2O2 burst was only observed in the wild-type ovules on the day of anthesis. Cotton boll injection experiments in combination with electron microscope observation collectively indicated that H2O2 burst, which is negatively regulated by ascorbate peroxidases (APx), plays an important role in the fiber initiation process. Taken together, our study demonstrates a putative network of DAP species related to fiber initiation in cotton ovules and provides a foundation for future studies on the specific functions of these proteins in fiber development.

Differential physiological, ultramorphological and metabolic responses of cotton cultivars under cadmium stress.

Cadmium (Cd) stress may cause serious physiological, ultramorphological and biochemical anomalies in plants. Cd-induced physiological, subcellular and metabolic alterations in two transgenic cotton cultivars (BR001, GK30) and their parent line (Coker 312) were evaluated using 10, 100 and 1000 µM Cd. Germination, fresh biomass of roots, stems and leaves were significantly inhibited at 1000 µM Cd. Root volume tolerance index significantly increased (124.16%) in Coker 312 at 1000 µM Cd. In non-Cd stressed conditions, electron micrographs showed well-configured root meristem and leaf mesophyll cells. At 1000 µM Cd, greater ultramorphological alterations were observed in BR001 followed by GK30 and Coker 312. These changes were observed in nucleus, vacuoles, mitochondria and chloroplast. Dense precipitates, probably Cd, were seen in vacuoles, which were also attached to the cell walls. A considerable increase in number of nuclei, vacuoles, starch granules and plastoglobuli was observed in the electron micrographs of both roots and leaves at 1000 µM Cd. MDA contents were higher in roots of BR001 at 1000 µM Cd. Mean values of SOD activity in leaves of both BR001 and GK30 at 1000 µM Cd significantly increased as compared to the controls. POD activity in roots of BR001 and Coker 312 was greater at all Cd (10, 100, 1000 µM) levels over the control. Regarding APX, highest percent increase (71.64%) in roots of GK30 at 1000 µM Cd was found. Non-significant differences in CAT activity were observed at all levels of Cd stress in leaves of BR001 and GK30. Both transgenic cotton cultivars and their parental line invariably responded towards Cd stress. However, Coker 312 showed Cd-resistant behavior as compared to its progeny lines (BR001 and GK30).

Cadmium-induced upregulation of lipid peroxidation and reactive oxygen species caused physiological, biochemical, and ultrastructural changes in upland cotton seedlings.

Cadmium (Cd) toxicity was investigated in cotton cultivar (ZMS-49) using physiological, ultrastructural, and biochemical parameters. Biomass-based tolerance index decreased, and water contents increased at 500 µM Cd. Photosynthetic efficiency determined by chlorophyll fluorescence and photosynthetic pigments declined under Cd stress. Cd contents were more in roots than shoots. A significant decrease in nutrient levels was found in roots and stem. A significant decrease in nutrient levels was found in roots and stems. In response to Cd stress, more MDA and ROS contents were produced in leaves than in other parts of the seedlings. Total soluble proteins were reduced in all parts except in roots at 500 µM Cd. Oxidative metabolism was higher in leaves than aerial parts of the plant. There were insignificant alterations in roots and leaves ultrastructures such as a little increase in nucleoli, vacuoles, starch granules, and plastoglobuli in Cd-imposed stressful conditions. Scanning micrographs at 500 µM Cd showed a reduced number of stomata as well as near absence of closed stomata. Cd depositions were located in cell wall, vacuoles, and intracellular spaces using TEM-EDX technology. Upregulation of oxidative metabolism, less ultrastructural modification, and Cd deposition in dead parts of cells show that ZMS-49 has genetic potential to resist Cd stress, which need to be explored.

Energy requirement for alkali assisted microwave and high pressure reactor pretreatments of cotton plant residue and its hydrolysis for fermentable sugar production for biofuel application.

In the present work, alkali assisted microwave pretreatment (AAMP) of cotton plant residue (CPR) with high pressure reactor pretreatment was compared. Further, modeling of AAMP was attempted. AAMP, followed by enzymatic saccharification was evaluated and the critical parameters were identified to be exposure time, particle size and enzyme loading. The levels of these parameters were optimized using response surface methodology (RSM) to enhance sugar yield. AAMP of CPR (1mm average size) for 6 min at 300 W yielded solid fractions that on hydrolysis resulted in maximum reducing sugar yield of 0.495 g/g. The energy required for AAMP at 300 W for 6 min was 108 kJ whereas high pressure pretreatment (180°C, 100 rpm for 45 min) requires 5 times more energy i.e., 540 kJ. Physiochemical characterization of native and pretreated feedstock revealed differences between high pressure pretreatment and AAMP.

Spectroscopic photoacoustic microscopy using a photonic crystal fiber supercontinuum source.

Photoacoustic microscopy (PAM) provides high resolution images with excellent image contrast based on optical absorption. The compact size and high repetition rate of pulsed microchip lasers make them attractive sources for PAM. However, their fixed wavelength output precludes their use in spectroscopic PAM. We are developing a tunable optical source based on a microchip laser that is suitable for spectroscopic PAM. Pulses from a 6.6 kHz repetition rate Q-switched Nd:YAG microchip laser are sent through a photonic crystal fiber with a zero dispersion wavelength at 1040 nm. The highly nonlinear optical propagation produces a supercontinuum spectrum spanning 500-1300 nm. A tunable band pass filter selects the desired wavelength band from the supercontinuum. Our PAM system employs optical focusing and a 25 MHz spherically focused detection transducer. En-face imaging experiments were performed at seven different wavelengths from 575 to 875 nm. A simple discriminant analysis of the multiwavelength photoacoustic data produces images that clearly distinguish the different absorbing regions of ink phantoms. These results suggest the potential of this compact tunable source for spectroscopic photoacoustic microscopy.

Shea meal and cotton stalk as potential fuels for co-combustion with coal.

The efficient management of waste biomass is an important environmental problem in agricultural countries. Often land-fill is the main disposal route with ramifications including CH(4) release having 21 times greater global warming potential per molecule than CO(2). Biomasses are considered to be CO(2)-neutral fuels when combusted. Moreover, they are renewable and covered by the renewable obligation scheme and eligible for certificates in the UK. The overall objective of the investigation is to assess the performance of selected biomass and coal co-firing under two different modes of operation, air-staging and fuel-staging with the benefit of reduced-NO(x) and SO(2) emissions in power plant. The biomasses chosen for the study, shea meal (SM) and cotton stalk (CS) have very different cellulose/lignin compositions and different reported thermal behaviour. A series of experiments have been carried out in a 20 kW, down fired combustor using coal, shea meal-coal and cotton stalk-coal blends under un-staged, air-staged and fuel-staged co-combustion configurations. For air-staging, an optimum value of primary zone stoichiometry SR(1)=0.9 was found. Keeping it fixed, the shea meal and cotton stalk content in the coal-biomass blends was set to 5%, 10% and 15% on thermal basis. NO reductions of 51% and 60% were achieved using SM and CS, respectively, with an optimum thermal biomass blending ratio (BBR) of 10%. The results obtained were compared with un-staged and air-staged results for coal without the addition of biomass. Similarly for fuel-staging, keeping the length of the reburn and burnout zone fixed, SM and CS were evaluated as reductive fuel using different reburn fuel fractions (R(ff)) of 5%, 10%, 15% and 20%. NO reductions of 83% and 84% were obtained with an optimum R(ff) of 15% with an optimum reburn zone stoichiometry of SR(2)=0.8 for both SM and CS, respectively. SO(2) reduction and char burnout efficiency were also evaluated. It was found that addition of biomass coupled with air and fuel-staging techniques reduced-NO(x) and SO(2) simultaneously while at the same time improving the char burnout efficiency.

Fusarium verticillioides (Saccardo) Nirenberg associated with hardlock of cotton.

Boll rots of cotton (Gossypium hirsutum L.) are common in the humid areas of the Southeastern US. One type of boll damage that may be differentiated from others is hardlock, with symptoms that include compression of the fibers within individual locules of mature, open cotton bolls without further obvious disintegration of the lint or damage to the carpel wall. The principal economic effect is that the boll's lint is unharvestable by mechanical cotton pickers. This disease is endemic to the Southeast and can cause severe yield losses up to 70% in some fields. Scanning electron microscopy images of fibers from hardlocked bolls showed flattened and twisted tissue compared to fibers from healthy bolls. Fusarium verticillioides (Saccardo) Nirenberg was the fungus most commonly isolated from seeds of developing cotton bolls. Flowers inoculated with F. verticillioides on the day of bloom by spraying a spore suspension onto the flowers developed significantly (P < 0.05) more hardlock symptoms compared to untreated controls. The infection process was analyzed using a F. verticillioides isolate tagged with green fluorescent protein (GFP). When it was applied to cotton flowers on the day of bloom, the GFP-tagged F. verticillioides strain was detected in the stigma and style by 2 days after bloom (DAB) and in developing seeds at 4, 6, 8, 10, 16, 20, 40, and 60 (open bolls) DAB. By 8 DAB, the GFP F. verticillioides was isolated from over 80% of developing seeds.

Histological and ultrastructural observation reveals significant cellular differences between Agrobacterium transformed embryogenic and non-embryogenic calli of cotton.

Over the past few decades genetic engineering has been applied to improve cotton breeding. Agrobacterium medicated transformation is nowadays widely used as an efficient approach to introduce exogenous genes into cotton for genetically modified organisms. However, it still needs to be improved for better transformation efficiency and higher embryogenic callus induction ratios. To research further the difference of mechanisms for morphogenesis between embryogenic callus and non-embryogenic callus, we carried out a systematical study on the histological and cellular ultrastructure of Agrobacterium transformed calli. Results showed that the embryogenic callus developed nodule-like structures, which were formed by small, tightly packed, hemispherical cells. The surface of some embryogenic callus was covered with a fibrilar-like structure named extracellular matrix. The cells of embryogenic calli had similar morphological characteristics. Organelles of embryogenic callus cells were located near the nucleus, and chloroplasts degraded to proplastid-like structures with some starch grains. In contrast, the non-embryogenic calli were covered by oval or sphere cells or small clusters of cells. It was observed that cells had vacuolation of cytoplasm and plastids with a well organized endomembrane system. This study aims to understand the mechanisms of embryogenic callus morphogenesis and to improve the efficiency of cotton transformation in future.

A specialized outer layer of the primary cell wall joins elongating cotton fibers into tissue-like bundles.

Cotton (Gossypium hirsutum) provides the world's dominant renewable textile fiber, and cotton fiber is valued as a research model because of its extensive elongation and secondary wall thickening. Previously, it was assumed that fibers elongated as individual cells. In contrast, observation by cryo-field emission-scanning electron microscopy of cotton fibers developing in situ within the boll demonstrated that fibers elongate within tissue-like bundles. These bundles were entrained by twisting fiber tips and consolidated by adhesion of a cotton fiber middle lamella (CFML). The fiber bundles consolidated via the CFML ultimately formed a packet of fiber around each seed, which helps explain how thousands of cotton fibers achieve their great length within a confined space. The cell wall nature of the CFML was characterized using transmission electron microscopy, including polymer epitope labeling. Toward the end of elongation, up-regulation occurred in gene expression and enzyme activities related to cell wall hydrolysis, and targeted breakdown of the CFML restored fiber individuality. At the same time, losses occurred in certain cell wall polymer epitopes (as revealed by comprehensive microarray polymer profiling) and sugars within noncellulosic matrix components (as revealed by gas chromatography-mass spectrometry analysis of derivatized neutral and acidic glycosyl residues). Broadly, these data show that adhesion modulated by an outer layer of the primary wall can coordinate the extensive growth of a large group of cells and illustrate dynamic changes in primary wall structure and composition occurring during the differentiation of one cell type that spends only part of its life as a tissue.

Cadmium-induced ultramorphological and physiological changes in leaves of two transgenic cotton cultivars and their wild relative.

The present study describes cadmium-induced alterations in the leaves as well as at the whole plant level in two transgenic cotton cultivars (BR001 and GK30) and their wild relative (Coker 312) using both ultramorphological and physiological indices. With elevated levels of Cd (i.e. 10, 100, 1000 microM), the mean lengths of root, stem and leaf and leaf width as well as their fresh and dry biomasses linearly decreased over their respective controls. Moreover, root, stem and leaf water absorption capacities progressively stimulated, which were high in leaves followed by roots and stems. BR001 accumulated more cadmium followed by GK30 and Coker 312. Root and shoot cadmium uptakes were significantly and directly correlated with each other as well as with leaf, stem and root water absorption capacities. The ultrastructural modifications in leaf mesophyll cells were triggered with increase in Cd stress regime. They were more obvious in BR001 followed by GK30 and Coker 312. Changes in morphology of chloroplast, increase in number and size of starch grains as well as increase in number of plastoglobuli were the noticed qualitative effects of Cd on photosynthetic organ. Cd in the form of electron dense granules could be seen inside the vacuoles and attached to the cell walls in all these cultivars. From the present experiment, it can be well established that both apoplastic and symplastic bindings are involved in Cd detoxification in these cultivars. Absence of tonoplast invagination reveals that Cd toxic levels did not cause water stress in any cultivars. Additionally, these cultivars possess differential capabilities towards Cd accumulation and its sequestration.

Cadmium-induced functional and ultrastructural alterations in roots of two transgenic cotton cultivars.

The toxic effect of cadmium (Cd) at increasing concentrations was studied with special attention being given to the root morphological and ultrastructural changes in two transgenic cotton cultivars viz. BR001 and GK30 and their wild relative viz. Coker 312. In comparison to their respective controls, low concentration (10 and 100microM) of Cd greatly stimulated seed germination, while it was inhibited by highest concentration of Cd (1000microM) in case of two transgenic cultivars. However, in Coker 312 the seed germination percentage progressively decreased over the control at all Cd levels. Various physiological and morphological parameters of the root and whole plant in both transgenic cotton cultivars and their relative wild cotton genotype respond differently towards the Cd toxicity. Bioavailability of Cd was concentration-dependent where seedling root captured more Cd as compared to shoot. BR001 accumulated more Cd followed by GK30, while Coker 312 was less Cd accumulator. The ultrastructural modifications in the root tip cells of both the transgenic cotton cultivars and their wild relative were also dose-dependent. With the increase in Cd levels, the fine structures of their root cells also invariably changed. Increase in plasmolysis of the plasma membrane, greater number of nucleoli and vacuoles and enlarged vacuoles could be observed in both transgenic cotton cultivars. In comparison to them, Coker 312 showed relatively well developed ultrastructures of the root tips except enlarged vacuoles and greater number of mitochondria. Moreover, the accumulation of Cd in the form of electron dense granules and crystals both in vacuoles and attached to cell walls were visible in both transgenic cotton cultivars and their wild relative. These results suggest that both transgenic cotton cultivars and their wild relative cotton genotype responded positively towards Cd stress at seedling stage, the internal Cd-detoxification might be through apoplastic and symplastic binding. Moreover, as a whole BR001 proved to be sensitive whereas; GK30 and Coker 312 were found as tolerant.