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Key Message: A long tracrRNA (tracr-L), which naturally act as single guide RNA, and its truncated version, Δtracr-L, from S. pyogenes, efficiently induce Cas9-mediated double-strand breaks (DSBs) in plant genomic loci, as demonstrated by in vitro cleavage assay and protoplast transfection. CRISPR-Cas system provides a form of immune memory in prokaryotes and archaea, protecting them against viruses and foreign genetic elements. In Streptococcus pyogenes, this system includes the pre-crRNA along with another non-coding RNA, tracrRNA, which aids in CRISPR-based immunity. In S. pyogenes, two distinct tracrRNAs are produced: a long form (tracr-L) and a short form (tracr-S). The tracr-S regulates crRNA biogenesis and Cas9 cleavage, while tracr-L suppresses CRISPR-Cas expression by targeting the Cas9 promoter to prevent autoimmunity. Deleting 79 nucleotides from tracr-L results in Δtracr-L, which retains similar functionality in gene repression. This study investigates, for the first time, the effectiveness of tracr-L, and Δtracr-L in genome editing within plant systems. In vitro cleavage assays using purified Cas9 and synthesized sgRNAs targeting the Cas9 gene, OsPDS, and the OsSWEET11 promoter revealed that across all target sites, tracr-S demonstrated the highest cleavage efficiency compared to tracr-L and Δtracr-L. For in vivo genome editing, we transfected rice protoplasts with tracr-L, Δtracr-L, and tracr-S, targeting three rice genes: OsPDS, OsSPL14, and the promoter of OsSWEET14. Amplicon deep sequencing revealed various types of indels at the target regions across all three tracrRNA versions, indicating comparable levels of efficiency. This study establishes the utility of both the long-form tracrRNA (tracr-L) and its truncated variant (Δtracr-L) in eukaryote genome editing. These two new forms of tracrRNA provide proof of concept and expand the CRISPR-Cas toolkit for plant genome editing applications, and for eukaryotes more broadly., Competing Interests: Declarations. Conflict of interest: The authors declare no conflict of interest., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Pigeon pea, vital for farmers in semi-arid regions, suffers yield losses from Fusarium wilt caused by Fusarium udum. This study demonstrates that introducing the rice oxalate oxidase 4 (Osoxo4) gene significantly boosts wilt resistance. Enhanced resistance in transgenic lines was confirmed through gene expression analysis, enzyme activity assays, biochemical assessments, histochemical staining and in vitro and in vivo bioassays, including spore germination tests. We performed proteomics and metabolomics analyses to investigate mechanisms of enhanced resistance. LC-MS/MS-based label-free proteomics of wilt-infected transgenic and wild-type pigeon pea leaves identified 2386 proteins, with 1048 showing significant abundance changes-738 upregulated and 310 downregulated-in transgenic plants. Notably, proteins such as HMG1/2-like protein, Putative nucleosome assembly protein C364.06, DEAD-box ATP-dependent RNA helicase 3, Lipoxygenase 1, Annexin D1 and Annexin-like protein RJ4 were significantly upregulated, indicating their potential role in developing wilt-resistant cultivars. Metabolomic analysis showed elevated levels of amino acids, sugars, oxalic acid, sugar alcohols and myo-inositol in transgenic pigeon pea, with upregulated pathways in Sugar and Starch Metabolism and Inositol Phosphate Metabolism, indicating enhanced resilience to wilt stress. This study highlights unique regulatory proteins and metabolites, offering insights into stress adaptation and guiding genetic interventions for breeding disease-resistant pigeon pea varieties., (© 2024 John Wiley & Sons Ltd.)

The CRISPR-Cas genome editing tools are revolutionizing agriculture and basic biology with their simplicity and precision ability to modify target genomic loci. Software-predicted guide RNAs (gRNAs) often fail to induce efficient cleavage at target loci. Many target loci are inaccessible due to complex chromatin structure. Currently, there is no suitable tool available to predict the architecture of genomic target sites and their accessibility. Hence, significant time and resources are spent on performing editing experiments with inefficient guides. Although in vitro-cleavage assay could provide a rough assessment of gRNA efficiency, it largely excludes the interference of native genomic context. Transient in-vivo testing gives a proper assessment of the cleavage ability of editing reagents in a native genomic context. Here, we developed a modified protocol that offers highly efficient protoplast isolation from rice, Arabidopsis, and chickpea, using a sucrose gradient, transfection using PEG (polyethylene glycol), and validation of single guide RNAs (sgRNAs) cleavage efficiency of CRISPR-Cas9. We have optimized various parameters for PEG-mediated protoplast transfection and achieved high transfection efficiency using our protocol in both monocots and dicots. We introduced plasmid vectors containing Cas9 and sgRNAs targeting genes in rice, Arabidopsis, and chickpea protoplasts. Using dual sgRNAs, our CRISPR-deletion strategy offers straightforward detection of genome editing success by simple agarose gel electrophoresis. Sanger sequencing of PCR products confirmed the editing efficiency of specific sgRNAs. Notably, we demonstrated that isolated protoplasts can be stored for up to 24/48 h with little loss of viability, allowing a pause between isolation and transfection. This high-efficiency protocol for protoplast isolation and transfection enables rapid (less than 7 days) validation of sgRNA cleavage efficiency before proceeding with stable transformation. The isolation and transfection method can also be utilized for rapid validation of editing strategies, evaluating diverse editing reagents, regenerating plants from transfected protoplasts, gene expression studies, protein localization and functional analysis, and other applications., Supplementary Information: The online version contains supplementary material available at 10.1007/s42994-024-00139-7., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© The Authors 2024.)

The estuaries provide the key pathway for travelling carbon across the land-ocean interfaces and behave as both source and sink of greenhouse gases (GHGs) in water-atmosphere systems. The sink-source characteristics of estuaries for GHGs vary spatially. The primary driving factors are adjacent ecologies (agriculture, aquaculture, etc.) and proximities to the sea. To study the sink-source characteristics of estuaries for GHGs (methane (CH 4 ), nitrous oxide (N 2 O) and carbon dioxide (CO 2 )), the water samples were collected from 53 different locations in the estuaries for estimation of dissolved GHGs concentration and air-water GHGs exchanges. The locations represent five zones (Zone I, II, III, IV and V) based on the type and degradation status of mangroves (degraded and undisturbed), anthropogenic activities, and distance from the sea. Zone I, III, V represents to the degraded mangroves far from sea, whereas, Zone II, IV surrounded by undisturbed mangroves and nearer to sea. The average dissolved CH 4 concentrations were higher in the estuaries which were adjacent to degraded mangroves (154.4 nmol L -1 ) than undisturbed mangroves (81.7 nmol L -1 ). Further, the average dissolved N 2 O concentrations were 48% higher in the estuaries nearer to degraded mangroves than that of undisturbed ones. Among the degraded mangrove sites, the dissolved CO 2 concentrations were higher at Zone I (30.1 μmol L -1 ) followed by Zone III and IV, whereas in undisturbed sites, it was higher in Zone IV (22.3 μmol L -1 ) than Zone II (17.6 μmol L -1 ). Among the 53 locations, 36, 51 and 33 locations acted as a sink (negative value of exchanges) for CH 4 , N 2 O and CO 2 , respectively. The higher sink potential for CH 4 was recorded to those estuaries adjacent to undisturbed mangroves (-791.69 μmol m -2  d -1 ) than the degraded ones (-23.18 μmol m -2  d -1 ). Similarly, the average air-water N 2 O and CO 2 exchanges were more negative in the estuaries which were nearer to undisturbed mangroves indicating higher sink potential. The pH, and salinity of the estuary water were negatively correlated with air-water CH 4 and N 2 O exchanges, whereas those were positively correlated with CO 2 exchanges. Significantly lower dissolved GHGs and air-water GHGs exchange was observed in the estuaries adjacent to the undisturbed mangrove as compared to the degraded mangrove. The reason behind higher sink behaviours of estuaries nearer to undisturbed mangroves are higher intrusion of seawater, less nutrient availability, higher salinity, low carbon contents and alkaline pH compared to estuaries adjacent to degraded mangroves and far from sea., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Low light (LL) stress during the grain-filling stage acutely impairs the quality and quantity of starch accumulation in rice grains. Here, we observed that LL-induced poor starch biosynthesis is modulated by auxin homeostasis, which regulates the activities of major carbohydrate metabolism enzymes such as starch synthase (SS) and ADP-glucose pyrophosphorylase (AGPase) in rice. Further, during the grain-filling period under LL, the starch/sucrose ratio increased in leaves but significantly decreased in the developing spikelets. This suggests poor sucrose biosynthesis in leaves and starch in the grains of the rice under LL. A lower grain starch was found to be correlated with the depleted AGPase and SS activities in the developing rice grains under LL. Further, under LL, the endogenous auxin (IAA) level in the spikelets was found to be synchronized with the expression of a heteromeric G protein gene, RGB1. Interestingly, under LL, the expression of OsYUC11 was significantly downregulated, which subsequently resulted in reduced IAA in the developing rice spikelets, followed by poor activation of grain-filling enzymes. This resulted in lowered grain starch accumulation, grain weight, panicle number, spikelet fertility, and eventually grain yield, which was notably higher in the LL-susceptible (GR4, IR8) than in the LL-tolerant (Purnendu, Swarnaprabha) rice genotypes. Therefore, we hypothesize that depletion in auxin biosynthesis under LL stress is associated with the downregulation of RBG1, which discourages the expression and activities of grain-filling enzymes, resulting in lower starch production, panicle formation, and grain yield in rice., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Rice is the major staple food crop for more than 50% of the world's total population, and its production is of immense importance for global food security. As a photophilic plant, its yield is governed by the quality and duration of light. Like all photosynthesizing plants, rice perceives the changes in the intensity of environmental light using phytochromes as photoreceptors, and it initiates a morphological response that is termed as the shade-avoidance response (SAR). Phytochromes (PHYs) are the most important photoreceptor family, and they are primarily responsible for the absorption of the red (R) and far-red (FR) spectra of light. In our endeavor, we identified the morphological differences between two contrasting cultivars of rice: IR-64 (low-light susceptible) and Swarnaprabha (low-light tolerant), and we observed the phenological differences in their growth in response to the reduced light conditions. In order to create genomic resources for low-light tolerant rice, we constructed a subgenomic library of Swarnaprabha that expedited our efforts to isolate light-responsive photoreceptors. The titer of the library was found to be 3.22 × 10 5 cfu/mL, and the constructed library comprised clones of 4-9 kb in length. The library was found to be highly efficient as per the number of recombinant clones. The subgenomic library will serve as a genomic resource for the Gramineae community to isolate photoreceptors and other genes from rice.

C 4 plants have the inherent capacity to concentrate atmospheric CO 2 in the vicinity of RuBisCo, thereby increasing carboxylation, and inhibiting photorespiration. Carbonic anhydrase (CA), the first enzyme of C 4 photosynthesis, converts atmospheric CO 2 to HCO 3 - , which is utilized by PEPC to produce C 4 acids. Bioengineering of C 4 traits into C 3 crops is an attractive strategy to increase photosynthesis and water use efficiency. In the present study, we isolated the PEPC gene from the C 4 plant Setaria italica and transferred it to C 3 rice. Overexpression of SiPEPC resulted in a 2-6-fold increment in PEPC enzyme activity in transgenic lines with respect to non-transformed control. Photosynthetic efficiency was enhanced in transformed plants, which was associated with increased ФPSII, ETR, lower NPQ, and higher chlorophyll accumulation. Water use efficiency was increased by 16-22% in PEPC transgenic rice lines. Increased PEPC activity enhanced quantum yield and carboxylation efficiency of PEPC transgenic lines. Transgenic plants exhibited higher light saturation photosynthesis rate and lower CO 2 compensation point, as compared to non-transformed control. An increase in net photosynthesis increased the yield by (23-28.9%) and biomass by (24.1-29%) in transgenic PEPC lines. Altogether, our findings indicate that overexpression of C 4- specific SiPEPC enzyme is able to enhance photosynthesis and related parameters in transgenic rice., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Masson SAS. All rights reserved.)

Genome editing technology has rapidly evolved to knock-out genes, create targeted genetic variation, install precise insertion/deletion and single nucleotide changes, and perform large-scale alteration. The flexible and multipurpose editing technologies have started playing a substantial role in the field of plant disease management. CRISPR-Cas has reduced many limitations of earlier technologies and emerged as a versatile toolbox for genome manipulation. This review summarizes the phenomenal progress of the use of the CRISPR toolkit in the field of plant pathology. CRISPR-Cas toolbox aids in the basic studies on host-pathogen interaction, in identifying virulence genes in pathogens, deciphering resistance and susceptibility factors in host plants, and engineering host genome for developing resistance. We extensively reviewed the successful genome editing applications for host plant resistance against a wide range of biotic factors, including viruses, fungi, oomycetes, bacteria, nematodes, insect pests, and parasitic plants. Recent use of CRISPR-Cas gene drive to suppress the population of pathogens and pests has also been discussed. Furthermore, we highlight exciting new uses of the CRISPR-Cas system as diagnostic tools, which rapidly detect pathogenic microorganism. This comprehensive yet concise review discusses innumerable strategies to reduce the burden of crop protection., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Low light intensity affects several physiological parameters during the different growth stages in rice. Plants have various regulatory mechanisms to cope with stresses. One of them is the differential and temporal expression of genes, which is governed by post-transcriptional gene expression regulation through endogenous miRNAs. To decipher low light stress-responsive miRNAs in rice, miRNA expression profiling was carried out using next-generation sequencing of low-light-tolerant (Swarnaprabha) and -sensitive (IR8) rice genotypes through Illumina sequencing. Swarnaprabha and IR8 were subjected to 25% low light treatment for one day, three days, and five days at the active tillering stage. More than 43 million raw reads and 9 million clean reads were identified in Swarnaprabha, while more than 41 million raw reads and 8.5 million clean reads were identified in IR8 after NGS. Importantly, 513 new miRNAs in rice were identified, whose targets were mostly regulated by the genes involved in photosynthesis and metabolic pathways. Additionally, 114 known miRNAs were also identified. Five novel ( osa-novmiR1 , osa-novmiR2 , osa-novmiR3 , osa-novmiR4 , and osa-novmiR5 ) and three known ( osa-miR166c-3p , osa-miR2102-3p , and osa-miR530-3p ) miRNAs were selected for their expression validation through miRNA-specific q RT-PCR. The expression analyses of most of the predicted targets of corresponding miRNAs show negative regulation. Hence, miRNAs modulated the expression of genes providing tolerance/susceptibility to low light stress. This information might be useful in the improvement of crop productivity under low light stress., Competing Interests: The authors declare that they have no conflict of interest.

Carbon concentrating mechanisms (CCMs) in plants are abaptive features that have evolved to sustain plant growth in unfavorable environments, especially at low atmospheric carbon levels and high temperatures. Uptake of CO 2 and its storage in the aerenchyma tissues of Lycopsids and diurnal acidity fluctuation in aquatic plants during the Palaeozoic era (ca. 300 Ma.) would represent the earliest evolution of a CCM. The CCM parts of the dark reactions of photosynthesis have evolved many times, while the light reactions are conserved across plant lineages. A C 4 type CCM, leaf C 4 photosynthesis is evolved in the PACMAD clade of the Poaceae family. The evolution of C 4 photosynthesis from C 3 photosynthesis was an abaptation. Photosynthesis in reproductive tissues of sorghum and maize (PACMAD clade) has been shown to be of a weaker C 4 type (high CO 2 compensation point, low carbon isotope discrimination, and lack of Rubisco compartmentalization, when compared to the normal C 4 types) than that in the leaves (normal C 4 type). However, this does not fit well with the character polarity concept from an evolutionary perspective. In a recent model proposed for CCM evolution, the development of a rudimentary CCM prior to the evolution of a more efficient CCM (features contrasting to a weaker C 4 type, leading to greater biomass production rate) has been suggested. An intermediate crassulacean acid metabolism (CAM) type of CCM (rudimentary) was reported in the genera, Brassia, Coryanthes, Eriopsis, Peristeria, of the orchids (well-known group of plants that display the CAM pathway). Similarly, we propose here the evolution of a rudimentary CCM (C 4 -like type pathway) in the non-foliar tissues of the Poaceae, prior to the evolution of the C 4 pathway as identified in the leaves of the C 4 species of the PACMAD clade., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Seed germination plays cardinal roles in seedling establishment and their successive growth. However, seed germination is retarded by far-red (FR) enrichment under low light stress, and the inhibitory signalling mechanism remains ambiguous. Our results indicated that low light treatment, both in the open and growth chamber conditions, inhibits rice seed germination by decreasing the gibberellin (GA) contents. To explore the mechanism of GA-deficiency under low light stress, differential expression profiling of GA-anabolic, -catabolic, ABA -anabolic, -catabolic, and SLR1 was investigated, revealing that expression of ABA- anabolic, GA-catabolic genes and SLR1 was upregulated with a simultaneous downregulation of ABA-catabolic and GA-anabolic genes under low light treatment. These results suggested that FR-induced GA inadequacy is resulted by upregulation of SLR1 and GA-catabolism genes consequently increase DELLA that further subsided GA-responses in the germinating rice seeds. Moreover, we provided evidence that FR-induced GA inadequacy demotes rice seed germination by decreasing amylase activity, eventually decreasing the carbohydrate solubilization in the germinating seeds. Finally, we suggest that under low light stress, due to a retarded conversion of phytochrome A to their bioactive form, the ABA-catabolic genes were eventually upregulated with a simultaneous downregulation of GA-anabolic genes. Consequently, a lower GA pool fails to leverage the GA-dependent DELLA degradation, further shutting down the expected GA responses that reduce germination efficiency under FR-enriched light., Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01167-7., Competing Interests: Conflict of interestThe authors did not receive support from any organization for the submitted work. The authors have no competing interests to declare that are relevant to the content of this article. There is no conflict of interest among the authors regarding the authorship of this manuscript., (© Prof. H.S. Srivastava Foundation for Science and Society 2022.)

Drought/osmotic stress is the single largest production constraint in rain-fed rice cultivation. Different members of the DREB gene family are known to contribute to osmotic stress tolerance. In this study, an attempt was made to understand their relative contribution towards osmotic stress tolerance in indica and japonica ecotypes of rice. Two genotypes (one tolerant and one susceptible) from each ecotype were grown hydroponically, and 21-day-old seedlings were subjected to polyethylene glycol-induced osmotic stress (15% PEG-6000, equivalent to -3.0 bars osmotic potential). The tolerant genotypes CR143 and Moroberekan were found to have superior root traits (total root length, surface area and volume), better plant water status and increased total dry biomass as compared to their susceptible counterparts after 10 days of osmotic stress. Different members of the DREB gene family were differentially induced in response to osmotic shock (1 h after stress) and osmotic stress (24 h after stress), which also differed between the two rice ecotypes. From the gene expression profiles of 10 DREB genes (both DREB1 and DREB2 families), in indica two DREB genes, DREB1B and DREB1G, were significantly correlated with stress tolerance indices, whereas in japonica significant correlations with five DREB genes (DREB1A, DREB1B, DREB1D, DREB1E and DREB2B) were observed. We found that only one member, i.e. DREB1B, showed a significant correlation with drought tolerance indices in both indica and japonica ecotypes. This study provides an overview of the relative contribution of different members of the DREB gene family and their association with drought/osmotic stress tolerance in rice., (© 2021 German Society for Plant Sciences, Royal Botanical Society of the Netherlands.)

Mangroves play a key role in ecosystem balancing and climate change mitigation. It acts as a source and sink of methane (CH 4 ), a major greenhouse gas responsible for climate change. Energy metabolic pathways of methane production (methanogenesis) and oxidation (methanotrophy) are directly driven by sulphur (S) and nitrogen (N) metabolism and salinity in coastal wetlands. To investigate, how mangrove-degradations, affect the source-sink behaviour of CH 4 ; the pathways of CH 4 , S and N were studied through whole-genome metagenomic approach. Soil samples were collected from degraded and undisturbed mangrove systems in Sundarban, India. Structural and functional microbial diversities (KEGG pathways) of CH 4 , S and N metabolism were analysed and correlated with labile carbon pools and physico-chemical properties of soil. Overall, the acetoclastic pathway of methanogenesis was dominant. However, the relative proportion of conversion of CO 2 to CH 4 was more in degraded mangroves. Methane oxidation was higher in undisturbed mangroves and the serine pathway was dominant. After serine, the ribulose monophosphate pathway of CH 4 oxidation was dominant in degraded mangrove, while the xylulose monophosphate pathway was dominant in undisturbed site as it is more tolerant to salinity and higher pH. The assimilatory pathway (AMP) of S-metabolism was dominant in both systems. But in AMP pathway, adenosine triphosphate sulfurylase enzyme reads were higher in degraded mangrove, while NADPH-sulfite reductase abundance was higher in undisturbed mangrove due to higher salinity, and pH. In N-metabolism, the denitrification pathway was predominant in degraded sites, whereas the dissimilatory nitrate reduction pathway was dominant in undisturbed mangroves. The relative ratios of sulphur reducing bacteria (SRB): methanogens were higher in degraded mangrove; however, methanotrophs:methanogens was higher in undisturbed mangrove indicated lower source and greater sink capacity of CH 4 in the system. Microbial manipulation in mangrove-rhizosphere for regulating major energy metabolic pathways of methane could open-up a new window of climate change mitigation in coastal wetlands., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Photorespiration accounts for 20-50 % reduction in grain yield in C 3 crops. The process is essential to remove 2-phosphoglycolate produced due to the oxygenation activity of the ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO) enzyme. Attempts were made to improve photosynthesis through enriched CO 2 concentration by installing numerous photorespiratory bypass modules in the chloroplast of several crops. In this study, we have introduced Escherichia coli glycolate catabolic pathway (ECGC) into rice chloroplast to bypass photorespiration partially (PB) or completely (FB). Five genes encoding glyoxylate carboligase (GCL), tartronic semialdehyde reductase (TSR), and three subunits of glycolate dehydrogenase (GDH) were introduced to get FB plants, whereas only the three subunits of GDH were introduced to get PB plants. Southern analysis confirmed stable integration of the transgenes and their expression was confirmed by RT-qPCR analysis in the T3 progenies. Both FB and PB transformed lines exhibited increased photosynthetic efficiency, biomass, and grain yield than wild type (WT) with empty vector control. The introduction of ECGC pathway favoured the carboxylase activity of RuBisCO while decreasing its oxygenase activity fostering the functioning of Calvin-Benson cycle and resulting in an increased carbon-assimilation that was manifested in their superior architecture and harvest index. These findings will support rice and related cereal crop breeding programs to increase yield under elevated temperature and arid conditions., (Copyright © 2021 Elsevier B.V. All rights reserved.)

C4 plants are superior to C3 plants in terms of productivity and limited photorespiration. PPDK (pyruvate orthophosphate dikinase) and NADP-ME (NADP-dependent malic enzyme) are two important photosynthetic C4-specific enzymes present in the mesophyll cells of C4 plants. To evaluate the effect of C4 enzymes in rice, we developed transgenic rice lines by separately introducing Setaria italica PPDK [SiPPDK] and S. italica ME [SiME] gene constructs under the control of the green tissue-specific maize PPDK promoter. Rice plant lines for both constructs were screened using the polymerase chain reaction (PCR), Southern hybridization, and expression analysis. The best transgenic plant lines for each case were selected for physiological and biochemical characterization. The results from qRT-PCR and enzyme activity analysis revealed higher expression and activity of both PPDK and NADP-ME genes compared with the nontransformed and empty-vector-transformed plants. The average photosynthetic efficiency of transgenic plant lines carrying the PPDK and NADP-ME genes increased by 18% and 12%, respectively, and was positively correlated with the increased accumulation of photosynthetic pigment. The decrease in Fv/Fm, increased electron transport rate (ETR), and increased photochemical quenching (qP) compared with nontransformed control plants suggest that transgenic rice plants transferred more absorbed light energy to photochemical reactions than wild-type plants. SiME-transgenic plants displayed reduced leaf malate content and superior performance under water deficit conditions. Interestingly, the transgenic plants showed yield enhancement by exhibiting increased plant height, panicle length, panicle weight and thousand grain weight. Overall, the exogenous foxtail millet C4 gene PPDK enhanced photosynthesis and yield to a greater extent than NADP-ME., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Base editors have drawn considerable academic and industrial attention in recent years because of their ability to alter single DNA bases with precision. However, the existing cytosine and adenine base editors can only install transition mutations. Three recent studies (Kurt et al.,Zhao et al., and Chen et al.) expand the base editing toolbox by developing cytosine transversion base editors., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Rice grain yield is drastically reduced under low light especially in kharif (wet) season due to cloudy weather during most part of crop growth. Therefore, 50-60% of yield penalty was observed. To overcome this problem, identification of low light tolerant rice genotypes with a high buffering capacity trait such as photosynthetic rate has to be developed. Sedoheptulose-1,7 bisphosphatase, a light-regulated enzyme, plays pivotal role in the Calvin cycle by regenerating the substrate (RuBP) for RuBisCo and therefore, indirectly regulates the influx of CO 2 for this crucial process. We found a potential role of SBPase expression and activity in low light tolerant and susceptible rice genotypes by analyzing its influence on net photosynthetic rate and biomass. We observed a significant relationship of yield with photosynthesis, SBPase expression and activity especially under low light conditions. Two tolerant and two susceptible rice genotypes were used for the present study. Tolerant genotypes exhibited significant but least reduction compared to susceptible genotypes in the expression and activity of SBPase, which was also manifested in its photosynthetic rate and finally in the grain yield under low light. However, susceptible genotypes showed significant reduction in SBPase activity along with photosynthesis and grain yield suggesting that tracking the expression and activity of SBPase could form a simple and reliable method to identify the low light tolerant rice cultivars. The data were analyzed using the Indostat 7.5, Tukey-Kramer method through Microsoft Excel 2019 and PAST4.0 software. The significant association of SBPase activity with the grain yield, net assimilation rate, electron transfer rate, biomass and grain weight were observed under low light stress. These traits should be considered while selecting and breeding for low light tolerant cultivars. Thus, SBPase plays a major role in the low light tolerance mechanism in rice., (© Prof. H.S. Srivastava Foundation for Science and Society 2020.)

Background: Phytic acid (PA) is an anti-nutrient present in cereals and pulses. It is known to reduce mineral bioavailability and inhibit starch-digesting α-amylase (which requires calcium for activity) in the human gut. In principle, the greater the amount of PA, the lower is the rate of starch hydrolysis. It is reflected in the lower glycemic index (GI) value of food. People leading sedentary lifestyles and consuming rice as a staple food are likely to develop type 2 diabetes. Hence, this study was planned to understand how PA content of different rice varieties affects the GI., Results: Rice Khira and Mugai which had very low PA (0.30 and 0.36 g kg -1 , respectively) had higher GI values and α-amylase activity, while Nua Dhusara and the pigmented rice Manipuri black rice (MBR) which had high PA (2.13 and 2.98 g kg -1 , respectively) showed low α-amylase activity and GI values. This relationship was statistically significant, though a weak relationship was found for the pigmented rice. Expression levels of MIPSI, IPKI and GBSSI markedly increased in the middle stage of grain development in all of the six genotypes having contrasting PA and GI. Maximum expression of MIPSI and IPKI was observed in Nua Dhusara and MBR (which had high PA) while that of GBSSI was observed in Khira and Mugai (with higher GI) at middle stage showing a negative correlation between PA and GI., Conclusions: The data indicate that high PA content in rice might have an adverse effect on starch digestibility resulting in slower starch digestion in the human gut and consequently low glycemic response. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Low light intensity is a great limitation for grain yield and quality in rice. However, yield is not significantly reduced in low light tolerant rice varieties. The work therefore planned for comparative transcriptome profiling under low light stress to decipher the genes involved and molecular mechanism of low light tolerance in rice. At active tillering stage, 50% low light exposure for 1 day, 3 days and 5 days were given to Swarnaprabha (low light tolerant) and IR8 (low light sensitive) rice varieties. Illumina (HiSeq) platform was used for transcriptome sequencing. A total of 6,652 and 12,042 genes were differentially expressed due to low light intensity in Swarnaprabha and IR8, respectively as compared to control. CAB, LRP, SBPase, MT15, TF PCL1 and Photosystem I & II complex related gene expressions were mostly increased in Swarnaprabha upon longer duration of low light exposure which was not found in IR8 as compared to control. Their expressions were validated by qRT-PCR. Overall study suggested that the maintenance of grain yield in the tolerant variety under low light might be results of accelerated expression of the genes which enable the plant to keep the photosynthetic processes moving at the same pace even under low light.

Photosynthetically active radiation (PAR) is one of the most important environmental factors that determine the productivity and grain quality of the crops. Continuous rainy days or cloudy weather throughout crop growth especially at critical stages often resulted in great loss of grain quality and yield in rice. Low light stress has rigorously constrained the rice production in various rice-growing regions, especially in Southeast Asia. Method and time of planting are the major management factors contributing to the higher yield potential of rice by influencing light harvesting and use efficiency. Present study was executed consecutively for 5 years (kharif seasons of 2012-2016) to determine whether planting time improves the radiation absorption and use efficiency in different duration rice cultivars. We evaluated the difference in plant growth and development leading to yield formation under different planting time which related to radiation incidence and interception. The results of the study revealed that PAR interception depends on morphological characters of cultivars and also with agronomic management such as transplanting time and method. Long duration cultivar intercepted more PAR but interception decreased due to late planting (3rd week of July), whereas short duration cultivars (Naveen) when planted earlier (1st week of June) could not effectively utilize intercepted PAR constraining the biomass accumulation and yield formation. Effect of planting density and crop architecture on PAR absorption was apparent among establishment methods as light interception at crop canopy was highest in the system of rice intensification and lowest in that of wet direct seeding. In general, Pooja as a long duration cultivar intercepted more PAR per day but when compared on same date of planting, the comparative absorption of radiation was 30.6% higher in Naveen. The lower yields in the wet season are attributed mostly to reduction in grain number per panicle or per unit land area, which is a consequence of high spikelet sterility. Grain yield of rice planted in July third week was reduced by 3.8, 12.3, and 6.9% over June first and third week and July first week, respectively, mainly due to spikelet sterility (26%) and lower grains per panicle (18%). Our results indicated that agronomic management like optimum time of sowing, cultivar duration, and establishment methods should be followed for yield improvement in tropical lowlands where light intensity is limiting due to prevailing weather situations.

Methane (CH 4 ) is predominantly produced in lowland rice soil, but its emission from soil to atmosphere primarily depends on passage/conduit or capillary pore spaces present in rice plants. The gas transport mechanism through aerenchyma pore spaces of rice cultivars was studied to explore the plant mediated CH 4 emission. Seven rice cultivars, based on the life cycle duration (LCD), were tested in tropical eastern India. Three LCD groups were, (a) Kalinga 1 and CR Dhan 204 (LCD: 110-120 days); (b) Lalat, Pooja and CR 1014 (LCD: 130-150 days); and (c) Durga and Varshadhan (LCD: 160-170 days). Rate of CH 4 emission, root exudates, root oxidase activities and shoot aerenchyma pore spaces were analyzed to study the mechanism of plant mediated emission from rice. Aerenchyma pore space was quantified in the hypothesis that it regulates the CH 4 transportation from soil to atmosphere. The ratio of pore space area to total space was lowest in Kalinga 1 cultivar (0.29) and highest was in Varshadhan (0.43). Significant variations in the methane emission were observed among the cultivars with an average emission rate ranged from 0.86 mg m -2  h -1 to 4.96 mg m -2  h -1 . The CH4 emission rates were lowest in short duration cultivars followed by medium and long duration ones. The greenhouse gas intensity considering average CH 4 emission rate per unit grain yield was also lowest (0.35) in Kalinga 1 and relatively less in short and medium duration cultivars. Root exudation was higher at panicle initiation (PI) than maximum tillering (MT) stage. Lowest exudation was noticed in (197.2 mg C plant -1  day -1 ) Kalinga 1 and highest in Varsadhan (231.7 mg C plant -1  day -1 ). So we can say, the rate of CH 4 emission was controlled by aerenchyma orientation, root exudation and biomass production rate which are the key specific traits of a cultivar. Identified traits were closely associated with duration and adaptability to cultivars grown in specific ecology. Therefore, there is possibility to breed rice cultivars depending on ecology, duration and having less CH 4 emission potential, which could be effectively used in greenhouse gas mitigation strategies., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The total digital information today amounts to 3.52 × 10 22 bits globally, and at its consistent exponential rate of growth is expected to reach 3 × 10 24 bits by 2040. Data storage density of silicon chips is limited, and magnetic tapes used to maintain large-scale permanent archives begin to deteriorate within 20 years. Since silicon has limited data storage ability and serious limitations, such as human health hazards and environmental pollution, researchers across the world are intently searching for an appropriate alternative. Deoxyribonucleic acid (DNA) is an appealing option for such a purpose due to its endurance, a higher degree of compaction, and similarity to the sequential code of 0's and 1's as found in a computer. This emerging field of DNA as means of data storage has the potential to transform science fiction into reality, wherein a device that can fit in our palms can accommodate the information of the entire world, as latest research has revealed that just four grams of DNA could store the annual global digital information. DNA has all the properties to supersede the conventional hard disk, as it is capable of retaining ten times more data, has a thousandfold storage density, and consumes 10 8 times less power to store a similar amount of data. Although DNA has an enormous potential as a data storage device of the future, multiple bottlenecks such as exorbitant costs, excruciatingly slow writing and reading mechanisms, and vulnerability to mutations or errors need to be resolved. In this review, we have critically analyzed the emergence of DNA as a molecular storage device for the future, its ability to address the future digital data crunch, potential challenges in achieving this objective, various current industrial initiatives, and major breakthroughs., Competing Interests: Compliance with ethical standardsThe authors declare that they have no competing interests.

Flooding is one of the major harmful abiotic stresses in the low lying areas of Asia and crop losses due to submergence are considerably high. Along with plant breeding techniques, agronomic management options in general and nutrient management in particular should be taken into consideration. Response of Sub 1 and non-Sub1 cultivars of rice to post-flood nitrogen (N) management under variable flood water was compared at maximum tillering stage. Submergence tolerance on survival, leaf senescence, metabolic changes, and anti-oxidant enzymatic activities were evaluated. Sub1 cultivars proved their superiority over IR-20 in terms of significantly higher survival, anti-oxidant enzymes and lower metabolic changes. Turbid water resulted in lower survival because of poor light transmission, chlorophyll retention and silt deposition. Basal phosphorus reduced the elongation, senescence and ethylene accumulation. Post-flood foliar spray of urea substantially increased the chlorophyll, soluble sugars and extenuated ethylene accumulation resulting in significantly higher survival. These nutrient management options can provide opportunities for better survival and productivity even under turbid water, helping farmers to cope with the existing problems in flood-prone areas.

An experiment was conducted to assess the effect of fly ash deposition without and with (0.25, 0.50, 1.0 and 1.5 g m(-2 )day(-1)) foliar dusting on the photosynthesis, stomatal conductance, transpiration, leaf temperature, albedo and productivity of rice. Dusting of 0.5 g m(-2 )day(-1) fly ash and above significantly reduced the photosynthesis, stomatal conductance, transpiration and albedo. Panicle initiation and flowering stages were more influenced by the fly ash deposition as compared to active tillering stage. At higher rates of fly ash deposition, all growth and yield parameters were significantly influenced due to increased heat load and reduced intercellular CO2 concentration. A significant reduction of 12.3, 15.7 and 20.2 % in grain yield was recorded over control when fly ash was dusted at 0.5, 1.0 and 1.5 g m(-2 )day(-1), respectively.

A field experiment was carried out to investigate the impact of elevated carbon dioxide (CO2) (CEC, 550 μmol mol(-1)) and elevated CO2+elevated air temperature (CECT, 550 μmol mol(-1) and 2°C more than control chamber (CC)) on soil labile carbon (C) and nitrogen (N) pools, microbial populations and enzymatic activities in relation to emissions of methane (CH4) and nitrous oxide (N2O) in a flooded alluvial soil planted with rice cv. Naveen in open top chambers (OTCs). The labile soil C pools, namely microbial biomass C, readily mineralizable C, water soluble carbohydrate C and potassium permanganate oxidizable C were increased by 27, 23, 38 and 37% respectively under CEC than CC (ambient CO2, 394 μmol mol(-1)). The total organic carbon (TOC) in root exudates was 28.9% higher under CEC than CC. The labile N fractions were also increased significantly (29%) in CEC than CC. Methanogens and denitrifier populations in rhizosphere were higher under CEC and CECT. As a result, CH4 and N2O-N emissions were enhanced by 26 and 24.6% respectively, under CEC in comparison to open field (UC, ambient CO2, 394 μmol mol(-1)) on seasonal basis. The global warming potential (GWP) was increased by 25% under CEC than CC. However, emissions per unit of grain yield under elevated CO2 and temperature were similar to those observed at ambient CO2. The stimulatory effect on CH4 and N2O emissions under CEC was linked with the increased amount of soil labile C, C rich root exudates, lowered Eh, higher Fe(+2) concentration and increased activities of methanogens and extracellular enzymes., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Plant height, biomass production, assimilatory functions and chlorophyll accumulation of Panicum maximum and Stylosanthes hamata in intercropping systems was influenced significantly under elevated CO2 (600 +/- 50 ppm) in open top chambers (OTCs). The plant height increased by 32.0 and 49.0% over the control in P. maximum and S. hamata respectively in intercropping system under elevated CO2 over open field grown crops (Ca). P. maximum and S. hamata produced 67 and 85% higher fresh and dry biomass respectively under elevated CO2. Rates of photosynthesis and stomatal conductance increased in both the crop species in intercropping systems under elevated CO2. The canopy photosynthesis (photosynthesis x leaf area index) of these crop species increased significantly under elevated CO2 over the open grown crops. The chlorophyll a and b accumulation were also higher in the leaves of both the crop species as grown in OTC with elevated CO2. The increased chlorophyll content, leaf area index and canopy photosynthesis led to higher growth and biomass production in these crop species under elevated CO2. The total carbon sequestration in crop biomass and soils during the three years was 21.53 Mg C/ha under elevated CO2. The data revealed that P. maximum and S. hamata intercropping system is the potential as a sink for the increasing level of CO2 in the atmosphere in the semi-arid tropics.

Seven species of genus Avena viz., Avena sativa, Avena strigosa, Avena brevis, Avena vaviloviana, Avena abyssinica, Avena marocana and Avena sterilis were used to study the impact of drought stress on lipid peroxidation and other antioxidant enzymes. Maximum increase in the catalase activity was recorded in A. vaviloviana (129.97%) followed by A. sativa (122.82%) and A. brevis (83.38%) at vegetative stage; however at flowering stage the maximum increase was reported in A. sativa (25.62%) followed by A. sterilis (20.46%) and A. brevis (18.53%). At vegetative stage drought, maximum increase in peroxidase activity was recorded in A. sativa (122.82%) followed by A. brevis (83.38%) and A. sterilis (49.78%). Flowering stage drought, showed maximum increase in A. Sativa (27.09%) followed by A. marocana (23.50%) and A. sterilis (20.46%). A. sativa and A. sterilis showed stress tolerance at both the stages by accumulating higher percentage of peroxidase followed by A. brevis at vegetative and A. marocana at flowering stage. Level of lipid peroxidation in terms of Malondialdehyde (MDA) content was increased in the leaves when plants were subjected to moisture stress. The rate of increase in lipid peroxidation occurs irrespective of stage however; maximum increase was recorded in A. strigosa at both the stages. Avena species which showed high level of MDA content, indicates more lipid peroxidation and more membrane permeability and are comparatively more susceptible for water stress than those which produce less Malondialdehyde (MDA) content at higher magnitude of water stress such species have better capability for moisture stress tolerance.