Combinative effect of pulsed-light irradiation and solid-state fermentation on ginkgolic acids, ginkgols, ginkgolides, bilobalide, flavonoids, product quality and sensory assessment of Ginkgo biloba dark tea.

Pulsed light (PL) is a prospective non-thermal technology that can improve the degradation of ginkgolic acid (GA) and retain the main bioactive compounds in Ginkgo biloba leaves (GBL). However, only using PL hasn't yet achieved the ideal effect of reducing GA. Fermentation of GBL to make ginkgo dark tea (GDT) could decrease GA. Because different microbial strains are used for fermentation, their metabolites and product quality might differ. However, there is no research on the combinative effect of PL irradiation fixation and microbial strain fermentation on main bioactive compounds and sensory assessment of GDT. In this research, first, Bacillus subtilis and Saccharomyces cerevisiae were selected as fermentation strains that can reduce GA from the five microbial strains. Next, the fresh GBL was irradiated by PL for 200 s (fluences of 0.52 J/cm2), followed by B. subtilis, S. cerevisiae, or natural fermentation to make GDT. The results showed that compared with the control (unirradiated and unfermented GBL) and the only PL irradiated GBL, the GA in GDT using PL + B. subtilis fermentation was the lowest, decreasing by 29.74%; PL + natural fermentation reduced by 24.53%. The total flavonoid content increased by 14.64% in GDT using PL + B. subtilis fermentation, whose phenolic and antioxidant levels also increased significantly. Sensory evaluation showed that the color, aroma, and taste of the tea infusion of PL + B. subtilis fermentation had the highest scores. In conclusion, the combined PL irradiation and solid-state fermentation using B. subtilis can effectively reduce GA and increase the main bioactive compounds, thus providing a new technological approach for GDT with lower GA.

A comprehensive assessment of photosynthetic acclimation to shade in C4 grass (Cynodon dactylon (L.) Pers.).

BACKGROUND: Light deficit in shaded environment critically impacts the growth and development of turf plants. Despite this fact, past research has predominantly concentrated on shade avoidance rather than shade tolerance. To address this, our study examined the photosynthetic adjustments of Bermudagrass when exposed to varying intensities of shade to gain an integrative understanding of the shade response of C4 turfgrass. RESULTS: We observed alterations in photosynthetic pigment-proteins, electron transport and its associated carbon and nitrogen assimilation, along with ROS-scavenging enzyme activity in shaded conditions. Mild shade enriched Chl b and LHC transcripts, while severe shade promoted Chl a, carotenoids and photosynthetic electron transfer beyond QA- (ET0/RC, φE0, Ψ0). The study also highlighted differential effects of shade on leaf and root components. For example, Soluble sugar content varied between leaves and roots as shade diminished SPS, SUT1 but upregulated BAM. Furthermore, we observed that shading decreased the transcriptional level of genes involving in nitrogen assimilation (e.g. NR) and SOD, POD, CAT enzyme activities in leaves, even though it increased in roots. CONCLUSIONS: As shade intensity increased, considerable changes were noted in light energy conversion and photosynthetic metabolism processes along the electron transport chain axis. Our study thus provides valuable theoretical groundwork for understanding how C4 grass acclimates to shade tolerance.

Assessment of heavy metals in soil, leaf litter, and their bioaccumulation in terrestrial macroinvertebrates in Sohag Governorate, Egypt.

The presence of elevated levels of heavy metals in soil poses a significant environmental concern with implications for human health and other organisms. The main objective of our study was to reduce the gap information of seasonal abundance, distribution of heavy metals in soil, leaf litter, and some macroinvertebrates in a citrus orchard (Citrus sinensis) in Sohag Governorate, Egypt. The heavy metals copper (Cu), zinc (Zn), lead (Pb), and cadmium (Cd) were determined by atomic absorption spectrometry. Degree of contamination (DC) was determined for both soil and leaf litter contamination. However, the bioaccumulation factor (BAF) was estimated to determine metal accumulation in the macroinvertebrates including earwigs Anisolabis maritima, chilopoda Scolopendra moristans, spider Dysdera crocata, and earthworm Aporrectodea caliginosa. The study area had clay-loam with varying organic matter, salinity, and pH levels. The degree of contamination varied among seasons, with the highest levels typically observed in autumn in both soil and leaf litter. The soil ranged from low contamination (1.82) to high contamination levels (4.4), while the leaf litter showed extremely high (30.03) to ultra-high (85.92) contamination levels. The mean ecological risk index results indicated that the sampling area had moderate ecological risk levels for Cd (44.3), Zn (42.17), and Pb (80.05), and extremely high levels for Cu (342.5). Heavy metal concentrations in the selected fauna were the highest in autumn, and the bioaccumulation factor varied among species and seasons with some species classified as e-concentrators, micro-concentrators, and macro-concentrators of certain heavy metals. Scolopendra moristans exhibited the highest mean metal concentrations (Cd, Pb, and Zn), while Aporrectodea caliginosa had the lowest. Thus, the differences in heavy metal concentrations found in different soil taxa highlight the significance of taxing a holistic understanding of feeding mechanisms into account when evaluating the potential risk for animals that consume invertebrates.

Effects of foliar selenium, biochar, and pig manure on cadmium accumulation in rice grains and assessment of health risk.

Addressing cadmium (Cd) contamination in agricultural lands is crucial, given its health implications and accumulation in crops. This study used pot experiments to evaluate the impact of foliar selenium spray (Se) (0.40 mM), corn straw biochar (1%), and pig manure (1%) on the growth of rice plants, the accumulation of Cd in rice grain, and to examine their influence on health risk indices associated with Cd exposure. The treatments were designated as follows: a control group without any amendment (CK), biochar (T1), pig manure (T2), Se (T3), Se and biochar (T4), Se and pig manure (T5), and Se along with biochar and pig manure (T6). Our results indicated that the treatments affected soil pH and redox potential and improved growth and the nitrogen and phosphorus content in rice plants. The soil-plant analysis development (SPAD) meter readings of leaves during the tillering stage indicated a 5.27%-15.86% increase in treatments T2 to T6 compared to CK. The flag leaves of T2 exhibited increases of 12.06%-38.94% for electrolyte leakage and an 82.61%-91.60% decline in SOD compared to treatments T3 to T6. Treatments T1 to T6 increased protein content; however, amylose content was significantly reduced in T6. Treatment T6 recorded the lowest Cd concentration in rice grains (0.018 mg/kg), while T2 recorded the highest (0.051 mg/kg). The CK treatment group showed a grain Cd content reduction of 29.30% compared to T2. The assessment of acceptable daily intake, hazard quotient, and carcinogenic risk revealed an ascending order as follows: T6 < T3 < T5 < T4 < T1 < CK < T2. In conclusion, the application of treatment T6 demonstrates the potential to lower oxidative stress, enhance production, reduce cancer risk, and ensure the safe cultivation of rice in environments affected by Cd contamination.

Interspecific variation in Rubisco CO2/O2 specificity along the leaf economic spectrum across 23 woody angiosperm plants in the Pacific islands.

The coordinated interspecific variation in leaf traits and leaf lifespan is known as the leaf economic spectrum (LES). The limitation of CO2 diffusion to chloroplasts within the lamina is significant in C3 photosynthesis, resulting in a shortage of CO2 for Rubisco. Although Rubisco CO2/O2 specificity (SC/O) should be adaptively adjusted in response to the interspecific variation in CO2 concentrations [CO2] associated with Rubisco, SC/O variations across species along the LES remain unknown. We investigated the coordination among leaf traits, including SC/O, CO2 conductance, leaf protein content, and leaf mass area, across 23 woody C3 species coexisting on an oceanic island through phylogenetic correlation analyses. A high SC/O indicates a high CO2 specificity of Rubisco. SC/O was negatively correlated with [CO2] at Rubisco and total CO2 conductance within lamina, while it was positively correlated with leaf protein across species, regardless of phylogenetic constraint. A simulation analysis shows that the optimal SC/O for maximizing photosynthesis depends on both [CO2] at Rubisco sites and leaf protein per unit leaf area. SC/O is a key parameter along the LES axis and is crucial for maximizing photosynthesis across species and the adaptation of woody plants.

Combined leaf gas-exchange system for model assessment.

Leaf gas-exchange measurements are useful in assessing plant environmental responses. However, uncertainties in the leaf gas-exchange model potentially limit its application. The main challenge in the model-dependent calculations is to detect violations of assumptions. Here, we developed a system that integrates into one instrument the direct measurement of leaf intercellular CO2 concentration and the standard open-flow (OF) and novel open-diffusion (OD) systems for flux measurement. In the OD system, a gas-permeable membrane between the leaf ambient air and outside air creates CO2 and H2O differentials, rather than the air flow in the OF chamber. We measured hypostomatous and amphistomatous leaves of several species with different photosynthetic capacities [sunflower (Helianthus annuus), grape (Vitis vinifera), lemon (Citrus limon), and cherry (Prunus avium)]. The CO2 and H2O differentials in the OD system strictly depend on the flux measured by the OF system. The lower permeability of the membrane resulted in a larger differential per flux, indicating that the OD system can increase the resolution for a small flux. An analysis of the conductance model along with observations suggested that cuticle and leaf intercellular conductances and the unsaturation of leaf humidity contributed to discrepancies between the direct measurement and standard calculation. The combined system developed here provides an opportunity to address these overlooked concepts in leaf gas exchange.

Contrasting coordination of non-structural carbohydrates with leaf and root economic strategies of alpine coniferous forests.

Non-structural carbohydrates (NSCs), as the labile fraction and dominant carbon currency, are essential mediators of plant adaptation to environments. However, whether and how NSC coordinates with plant economic strategy frameworks, particularly the well-recognized leaf economics spectrums (LES) and root economics space (RES), remains unclear. We examined the relationships between NSC and key plant economics traits in leaves and fine roots across 90 alpine coniferous populations on the Tibetan Plateau, China. We observed contrasting coordination of NSC with economics traits in leaves and roots. Leaf total NSC and soluble sugar aligned with the leaf economic spectrum, conveying a trade-off between growth and storage in leaves. However, NSC in roots was independent of the root economic spectrum, but highly coordinated with root foraging, with more starch and less sugar in forage-efficient, thinner roots. Further, NSC-trait coordination in leaves and roots was, respectively, driven by local temperature and precipitation. These findings highlight distinct roles of NSC in shaping the above- and belowground multidimensional economics trait space, and NSC-based carbon economics provides a mechanistic understanding of how plants adapt to heterogeneous habitats and respond to environmental changes.

Bioaccumulation, transfer characteristics of metals in six vascular plants, and soil pollution assessment from Wachangping karst bauxite residue areas.

Bauxite residue (BR) is a large volume by-product generated during bauxite smelting process and metal pollution problem is becoming increasingly prominent in residue areas. Accumulation and transfer of metals in six vascular plants were analyzed and soil environment was evaluated. Results found levels of Al (2,110-26,280 mg kg-1), Fe (990 to 9,880 mg kg-1), Ca (8,020 to 49,250 mg kg-1), Mg (2,060 to 17,190 mg kg-1), K (16,840 to 39,670 mg kg-1), and Ti (80 to 1,240 mg kg-1) in plants. Metal concentrations in soils exceeded background levels. Bioconcentration factor (BCF) found that Al, Fe, and Ti in plants (roots, stems, and leaves) were relatively depleted (BCF <1). Transfer factor (TF) of Al, Fe, Ca, K, and Ti in plants was distinctly higher than 1 and mainly concentrated in stems and leaves. Pollution indices revealed that soil environment was at moderated to serious contaminated risk. Principal components analysis (PCA) showed that Artemisia caruifolia Buch. and Siegesbeckia orientalis L. plants had a good ability to absorb Al and Fe, which can be used as biological indicators and restoration materials.

Currently, soil environment was exposed to moderated to serious contaminated risk from Wachangping karst bauxite residue areas.Bioconcentration factor (BCF) analysis found that Al, Fe, and Ti in six vascular plants (roots, stems, and leaves) were relatively depleted (BCF <1).Transfer factor (TF) of Al, Fe, Ca, K, and Ti in vascular plants was distinctly higher than 1, which mainly concentrated in stems and leaves.PCA revealed that Artemisia caruifolia Buch. and Siegesbeckia orientalis L. plants had a good ability to absorb Al and Fe, which can be used as biological indicators and ecological restoration materials.

Leaf phosphorus fractions vary with leaf economic traits among 35 Australian woody species.

Adaptations of plants to phosphorus (P) deficiency include reduced investment of leaf P in storage (orthophosphates in vacuoles), nucleic acids and membrane lipids. Yet, it is unclear how these adaptations are associated with plant ecological strategies. Five leaf P fractions (orthophosphate P, Pi ; metabolite P, PM ; nucleic acid P, PN ; lipid P, PL ; and residual P, PR ) were analysed alongside leaf economic traits among 35 Australian woody species from three habitats: one a high-P basalt-derived soil and two low-P sandstone-derived soils, one undisturbed and one disturbed by human activities with artificial P inputs. Species at the undisturbed low-P site generally exhibited lower concentrations of total leaf P ([Ptotal ]), primarily associated with lower concentrations of Pi , and PN . The relative allocation of P to each fraction varied little among sites, except that higher PL per [Ptotal ] (rPL ) was recorded at the undisturbed low-P site than at the high-P site. This higher rPL , reflecting relative allocation to membranes, was primarily associated with lower concentrations of leaf nitrogen at the undisturbed low-P site than at the high-P site. Associations between leaf P fractions and leaf nitrogen may provide a basis for understanding the variation in plant ecological strategies dependent on soil P availability.

A comparative full-length transcriptomic resource provides insight into the perennial monocarpic mass flowering.

Perennial monocarpic mass flowering represents as a key developmental innovation in flowering time diversity in several biological and economical essential families, such as the woody bamboos and the shrubby Strobilanthes. However, molecular and genetic mechanisms underlying this important biodiversity remain poorly investigated. Here, we generated a full-length transcriptome resource incorporated into the BlueOmics database (http://blueomics.iflora.cn) for two Strobilanthes species, which feature contrasting flowering time behaviors. Using about 112 and 104 Gb Iso-seq reads together with ~185 and ~75 Gb strand-specific RNA seq data, we annotated 80 971 and 79 985 non-redundant full-length transcripts for the perennial polycarpic Strobilanthes tetrasperma and the perennial monocarpic Strobilanthes biocullata, respectively. In S. tetrasperma, we identified 8794 transcripts showing spatiotemporal expression in nine tissues. In leaves and shoot apical meristems at two developmental stages, 977 and 1121 transcripts were differentially accumulated in S. tetrasperma and S. biocullata, respectively. Interestingly, among the 33 transcription factors showing differential expression in S. tetrasperma but without differential expression in S. biocullata, three were involved potentially in the photoperiod and circadian-clock pathway of flowering time regulation (FAR1 RELATED SEQUENCE 12, FRS12; NUCLEAR FACTOR Y A1, NFYA1; PSEUDO-RESPONSE REGULATOR 5, PRR5), hence provides an important clue in deciphering the flowering diversity mechanisms. Our data serve as a key resource for further dissection of molecular and genetic mechanisms underpinning key biological innovations, here, the perennial monocarpic mass flowering.

Triose phosphate utilization in leaves is modulated by whole-plant sink-source ratios and nitrogen budgets in rice.

Triose phosphate utilization (TPU) is a biochemical process indicating carbon sink-source (im)balance within leaves. When TPU limits leaf photosynthesis, photorespiration-associated amino acid exports probably provide an additional carbon outlet and increase leaf CO2 uptake. However, whether TPU is modulated by whole-plant sink-source relations and nitrogen (N) budgets remains unclear. We address this question by model analyses of gas-exchange data measured on leaves at three growth stages of rice plants grown at two N levels. Sink-source ratio was manipulated by panicle pruning, by using yellower-leaf variant genotypes, and by measuring photosynthesis on adaxial and abaxial leaf sides. Across all these treatments, higher leaf N content resulted in the occurrence of TPU limitation at lower intercellular CO2 concentrations. Photorespiration-associated amino acid export was greater in high-N leaves, but was smaller in yellower-leaf genotypes, panicle-pruned plants, and for abaxial measurement. The feedback inhibition of panicle pruning on rates of TPU was not always observed, presumably because panicle pruning blocked N remobilization from leaves to grains and the increased leaf N content masked feedback inhibition. The leaf-level TPU limitation was thus modulated by whole-plant sink-source relations and N budgets during rice grain filling, suggesting a close link between within-leaf and whole-plant sink limitations.

Rethinking economic theories of plant water use.

A central assumption in plant ecophysiology is that carbon is the primary currency for plant fitness. To this end, plants are thought to maximize carbon gain and any deviations from maximum carbon gain are ascribed to resource limitations (e.g., temperature, drought), biophysical limitations (e.g., biophysical limits on cell size), or variation in plant life history that may prioritize future carbon gain over current carbon gain (i.e., applying an economic discount rate to carbon). Compared to living in water, living on land made accessing CO2 substantially easier: CO2 diffuses approximately 10,000 times faster in air than in water. However, because this CO2 must diffuse into the aqueous environment of the living mesophyll cells where photosynthetic metabolism occurs (The´roux-Rancourt et al. 2021), the greater CO2 supply of the terrestrial lifestyle also comes with a cost: losing approximately 200-400 molecules of water by transpiration for every molecule of CO2 fixed by photosynthesis (Nobel et al. 2005). Water, therefore, is considered a valuable resource to be conserved and not wasted. As such, much of the field of plant ecophysiology posits carbon as the central currency for which water is traded.

Assessment of Salt Stress to Arabidopsis Based on the Detection of Hydrogen Peroxide Released by Leaves Using an Electrochemical Sensor.

Salt stress will have a serious inhibitory effect on various metabolic processes of plant cells, this will lead to the excessive accumulation of reactive oxygen species (ROS). Hydrogen peroxide (H2O2) is a type of ROS that can severely damage plant cells in large amounts. Existing methods for assessing the content of H2O2 released from leaves under salt stress will cause irreversible damage to plant leaves and are unable to detect H2O2 production in real time. In this study, on the strength of a series of physiological indicators to verify the occurrence of salt stress, an electrochemical sensor for the detection of H2O2 released from leaves under salt stress was constructed. The sensor was prepared by using multi-walled carbon nanotube-titanium carbide-palladium (MWCNT-Ti3C2Tx-Pd) nanocomposite as substrate material and showed a linear response to H2O2 detection in the range 0.05-18 mM with a detection limit of 3.83 µM. Moreover, we measured the determination of H2O2 released from Arabidopsis leaves at different times of salt stress by the sensor, which was consistent with conventional method. This study demonstrates that electrochemical sensing is a desirable technology for the dynamic determination of H2O2 released by leaves and the assessment of salt stress to plants.

Impaired cell growth under ammonium stress explained by modeling the energy cost of vacuole expansion in tomato leaves.

Ammonium (NH4 + )-based fertilization efficiently mitigates the adverse effects of nitrogen fertilization on the environment. However, high concentrations of soil NH4 + provoke growth inhibition, partly caused by the reduction of cell enlargement and associated with modifications of cell composition, such as an increase of sugars and a decrease in organic acids. Cell expansion depends largely on the osmotic-driven enlargement of the vacuole. However, the involvement of subcellular compartmentation in the adaptation of plants to ammonium nutrition has received little attention, until now. To investigate this, tomato (Solanum lycopersicum) plants were cultivated under nitrate and ammonium nutrition and the fourth leaf was harvested at seven developmental stages. The vacuolar expansion was monitored and metabolites and inorganic ion contents, together with intracellular pH, were determined. A data-constrained model was constructed to estimate subcellular concentrations of major metabolites and ions. It was first validated at the three latter developmental stages by comparison with subcellular concentrations obtained experimentally using non-aqueous fractionation. Then, the model was used to estimate the subcellular concentrations at the seven developmental stages and the net vacuolar uptake of solutes along the developmental series. Our results showed ammonium nutrition provokes an acidification of the vacuole and a reduction in the flux of solutes into the vacuoles. Overall, analysis of the subcellular compartmentation reveals a mechanism behind leaf growth inhibition under ammonium stress linked to the higher energy cost of vacuole expansion, as a result of alterations in pH, the inhibition of glycolysis routes and the depletion of organic acids.

Comparative transcriptomics of tropical woody plants supports fast and furious strategy along the leaf economics spectrum in lianas.

Lianas, climbing woody plants, influence the structure and function of tropical forests. Climbing traits have evolved multiple times, including ancestral groups such as gymnosperms and pteridophytes, but the genetic basis of the liana strategy is largely unknown. Here, we use a comparative transcriptomic approach for 47 tropical plant species, including ten lianas of diverse taxonomic origins, to identify genes that are consistently expressed or downregulated only in lianas. Our comparative analysis of full-length transcripts enabled the identification of a core interactomic network common to lianas. Sets of transcripts identified from our analysis reveal features related to functional traits pertinent to leaf economics spectrum in lianas, include upregulation of genes controlling epidermal cuticular properties, cell wall remodeling, carbon concentrating mechanism, cell cycle progression, DNA repair and a large suit of downregulated transcription factors and enzymes involved in ABA-mediated stress response as well as lignin and suberin synthesis. All together, these genes are known to be significant in shaping plant morphologies through responses such as gravitropism, phyllotaxy and shade avoidance.

The energy cost of repairing photoinactivated photosystem II: an experimental determination in cotton leaf discs.

Photosystem II (PSII), which splits water molecules at minimal excess photochemical potential, is inevitably photoinactivated during photosynthesis, resulting in compromised photosynthetic efficiency unless it is repaired. The energy cost of PSII repair is currently uncertain, despite attempts to calculate it. We experimentally determined the energy cost of repairing each photoinactivated PSII in cotton (Gossypium hirsutum) leaves, which are capable of repairing PSII in darkness. As an upper limit, 24 000 adenosine triphosphate (ATP) molecules (including any guanosine triphosphate synthesized at the expense of ATP) were required to repair one entire PSII complex. Further, over a 7-h illumination period at 526-1953 µmol photons m-2 s-1 , the ATP requirement for PSII repair was on average up to 4.6% of the ATP required for the gross carbon assimilation. Each of these two measures of ATP requirement for PSII repair is two- to three-fold greater than the respective reported calculated value. Possible additional energy sinks in the PSII repair cycle are discussed.

Assessment of the Photosynthetic Apparatus Functions by Chlorophyll Fluorescence and P700 Absorbance in C3 and C4 Plants under Physiological Conditions and under Salt Stress.

Functions of the photosynthetic apparatus of C3 (Pisum sativum L.) and C4 (Zea mays L.) plants under physiological conditions and after treatment with different NaCl concentrations (0-200 mM) were investigated using chlorophyll a fluorescence (pulse-amplitude-modulated (PAM) and JIP test) and P700 photooxidation measurement. Data revealed lower density of the photosynthetic structures (RC/CSo), larger relative size of the plastoquinone (PQ) pool (N) and higher electron transport capacity and photosynthetic rate (parameter RFd) in C4 than in C3 plants. Furthermore, the differences were observed between the two studied species in the parameters characterizing the possibility of reduction in the photosystem (PSI) end acceptors (REo/RC, REo/CSo and δRo). Data revealed that NaCl treatment caused a decrease in the density of the photosynthetic structures and relative size of the PQ pool as well as decrease in the electron transport to the PSI end electron acceptors and the probability of their reduction as well as an increase in the thermal dissipation. The effects were stronger in pea than in maize. The enhanced energy losses after high salt treatment in maize were mainly from the increase in the regulated energy losses (ΦNPQ), while in pea from the increase in non-regulated energy losses (ΦNO). The reduction in the electron transport from QA to the PSI end electron acceptors influenced PSI activity. Analysis of the P700 photooxidation and its decay kinetics revealed an influence of two PSI populations in pea after treatment with 150 mM and 200 mM NaCl, while in maize the negligible changes were registered only at 200 mM NaCl. The experimental results clearly show less salt tolerance of pea than maize.

The carbon economics of vegetative phase change.

Across plant species and biomes, a conserved set of leaf traits govern the economic strategy used to assimilate and invest carbon. As plants age, they face new challenges that may require shifts in this leaf economic strategy. In this study, we investigate the role of the developmental transition, vegetative phase change (VPC), in altering carbon economics as plants age. We used overexpression of microRNA 156 (miR156), the master regulator of VPC, to modulate the timing of VPC in Populus tremula x alba, Arabidopsis thaliana and Zea mays to understand the impact of this transition on leaf economic traits, including construction cost, payback time and return on investment. Here, we find that VPC causes a shift from a low-cost, quick return juvenile strategy to a high-cost, high-return adult strategy. The juvenile strategy is advantageous in light-limited conditions, whereas the adult strategy provides greater returns in high light. The transition between these strategies is correlated with the developmental decline in the level of miR156, suggesting that is regulated by the miR156/SPL pathway. Our results provide an ecophysiological explanation for the existence of juvenile and adult leaf types and suggest that natural selection for these alternative economic strategies could be an important factor in plant evolution.

Engineered resistance and risk assessment associated with insecticidal and weeds resistant transgenic cotton using wister rat model.

Stacking multiple genes into cotton crop to cop up multiple biotic stresses such as insects and weeds is a promising tool to save crop from losses. Transgenic cotton variety, VH-289, with double Bt and cp4EPSPS genes under the control of 35S promoter was used for the expression analyses and biosafety studies. The transgenic cotton plants were screened through PCR amplification of fragments, 1.7 kb for Cry1Ac, 582 bp for Cry2A and 250 bp for cp4EPSPS; which confirmed the presence of all genes transformed in transgenic cotton. The Cry1Ac + Cry2A and cp4EPSPS proteins were quantified through ELISA in transgenic cotton plants. The Glyphosate assay performed by spraying 1900 mL per acre of glyphosate Roundup further confirmed complete survival of transgenic cotton plants as compared to the non-transgenic cotton plants and all weeds. Similarly, insect infestation data determined that almost 99% insect mortality was observed in controlled field grown transgenic cotton plants as compared to the non-transgenic control plants. Evaluation of effect of temperature and soil nutrients availability on transgene expression in cotton plants was done at two different cotton growing regions, Multan and Lahore, Pakistan and results suggested that despite of higher temperature in Multan field, an increased level of Cry and cp4EPSPS proteins was recorded due to higher soil organic matter availability compared to Lahore field. Before commercialization of any transgenic variety its biosafety study is mandatory so, a 90 days biosafety study of the transgenic cotton plants with 40% transgenic cottonseeds in standard diet showed no harmful effect on wister rat model when studied for liver function, renal function and serum electrolyte.

Assessment of Protein Synthesis in Mitochondria Isolated from Rosette Leaves and Liquid Culture Seedlings of Arabidopsis.

Mitochondria are subcellular organelles with their own genome and expression system, including translation machinery to make proteins. Several independent studies have shown that translation is an essential regulatory step in expression of the plant mitochondrial genome. Thus, the study of mitochondrial translation seems to be crucial for the comprehension of plant mitochondrial biogenesis and maintenance. In organello protein synthesis in isolated mitochondria is a direct method to visualize the translational products of this organellar genetic system. In this method, highly purified, functional mitochondria synthesize proteins in the presence of radiolabeled amino acids, such as methionine, and an energy regeneration system. The labeled, newly synthesized polypeptides are separated by SDS-polyacrylamide gel electrophoresis and are detected by autoradiography. Here we describe the detailed protocol for in organello labeling of translation products that was optimized for mitochondria isolated from rosette leaves and liquid culture seedlings of Arabidopsis thaliana plants.