The Arabidopsis BUB1/MAD3 family protein BMF3 requires BUB3.3 to recruit CDC20 to kinetochores in spindle assembly checkpoint signaling.

The spindle assembly checkpoint (SAC) ensures faithful chromosome segregation during cell division by monitoring kinetochore-microtubule attachment. Plants produce both sequence-conserved and diverged SAC components, and it has been largely unknown how SAC activation leads to the assembly of these proteins at unattached kinetochores to prevent cells from entering anaphase. In Arabidopsis thaliana, the noncanonical BUB3.3 protein was detected at kinetochores throughout mitosis, unlike MAD1 and the plant-specific BUB1/MAD3 family protein BMF3 that associated with unattached chromosomes only. When BUB3.3 was lost by a genetic mutation, mitotic cells often entered anaphase with misaligned chromosomes and presented lagging chromosomes after they were challenged by low doses of the microtubule depolymerizing agent oryzalin, resulting in the formation of micronuclei. Surprisingly, BUB3.3 was not required for the kinetochore localization of other SAC proteins or vice versa. Instead, BUB3.3 specifically bound to BMF3 through two internal repeat motifs that were not required for BMF3 kinetochore localization. This interaction enabled BMF3 to recruit CDC20, a downstream SAC target, to unattached kinetochores. Taken together, our findings demonstrate that plant SAC utilizes unconventional protein interactions for arresting mitosis, with BUB3.3 directing BMF3's role in CDC20 recruitment, rather than the recruitment of BUB1/MAD3 proteins observed in fungi and animals. This distinct mechanism highlights how plants adapted divergent versions of conserved cell cycle machinery to achieve specialized SAC control.

Transcriptome analysis with different leaf blades identifies the phloem-specific phosphate transporter OsPHO1;3 required for phosphate homeostasis in rice.

Phosphate (Pi) is essential for plant growth and development. One strategy to improve Pi use efficiency is to enhance Pi remobilization among leaves. Using transcriptome analysis with first (top) and fourth (down) leaf blades from rice (Oryza sativa) in Pi-sufficient and deficient conditions, we identified 1384 genes differentially expressed among these leaf blades. These genes were involved in physiological processes, metabolism, transport, and photosynthesis. Moreover, we identified the Pi efflux transporter gene, OsPHO1;3, responding to Pi-supplied conditions among these leaf blades. OsPHO1;3 is highly expressed in companion cells of phloem, but not xylem, in leaf blades and induced by Pi starvation. Mutation of OsPHO1;3 led to Pi accumulation in second to fourth leaves under Pi-sufficient conditions, but enhanced Pi levels in first leaves under Pi-deficient conditions. These Pi accumulations in leaves of Ospho1;3 mutants resulted from induction of OsPHT1;2 and OsPHT1;8 in root and reduction of Pi remobilization in leaf blades, revealed by the decreased Pi in phloem of leaves. Importantly, lack of OsPHO1;3 caused growth defects under a range of Pi-supplied conditions. These results demonstrate that Pi remobilization is essential for Pi homeostasis and plant growth irrespective of Pi-supplied conditions, and OsPHO1;3 plays an essential role in Pi remobilization for normal plant growth.

A Phenomenological Investigation Into the Meaning of Food in Palliative Care Patients With Anorexia.

Little has been published on the meaning of food to palliative care patients with anorexia. Our study aims to investigate the meaning of food in palliative patients with anorexia. Fifteen patients with anorexia were recruited from the Palliative Care Unit of an acute hospital in Singapore from August 2018 to August 2021. A phenomenological methodology was employed to study the lived experience of anorexia and the meaning of food to palliative care patients. Our study findings revealed that food has social, physical, and emotional meaning in palliative care patients with anorexia. The social meaning of food was the predominant theme. Food was viewed as an important tool to bond and connect with their loved ones. It was perceived to be more important than the food itself and the taste of food was enhanced through social interactions. Food intake was related to physical strength and health status. Patients regarded eating as a way to improve their health status. Emotionally, eating was associated with positive feelings like enjoyment and freedom. Half of our participants felt that anorexia contributed to their low mood. Therefore, unlike the traditional focus of modifying the taste and quality of food in patients with anorexia, the authors recommended a focused assessment and management of the social aspect of anorexia on individual. This is important to mitigate the negative impact of anorexia, thus improving the quality of life and increasing their dignity towards the end of their lives.

Combined metabolomic and transcriptomic analysis evidences the interaction between sugars and phosphate in rice.

Phosphorus is one of the macro-elements required by plants, but phosphate (Pi), the only form that can be absorbed by plants, is always limited for plant growth and development. To adapt to Pi deficiency, plants have evolved a complex regulatory system to improve Pi acquisition and utilization efficiency. In this study, metabolomic and transcriptomic analyses were performed to exam the global metabolites and gene expressions profiles responding to Pi deficiency in rice. A total of 23 metabolites were co-changed in leaves and roots after Pi deficiency, with sucrose, trehalose and melibiose significant accumulated. A total of 779 genes were co-changed in these leaves and roots. Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that differentially expressed genes and differentially accumulated metabolites were co-enriched in galactose metabolism. Further exogenous sugars supply with rice roots could induce Pi starvation responsiveness and the expression of OsPHR2, which codes the central regulator for Pi starvation responsiveness in rice. This work revealed the interaction between sugars and phosphate in rice, and the importance of OsPHR2 in this interaction.

The differences and overlaps in the seed-resident microbiome of four Leguminous and three Gramineous forages.

Given the important roles that seed-borne endophytes can play on their plant hosts, comprehensive studies of the bacterial and fungal communities of seeds are of great importance. In this study, we assessed the seed endophytes of three gramineous (Avena sativa, Elymus sibiricus and Elymus dahuricus) and four leguminous (Vicia villosa, Trifolium repens, Trifolium pretense and Medicago sativa) forages using high-throughput sequencing. In total, 1013 distinct bacterial operational taxonomic units (OTUs) and 922 fungal OTUs were detected, with bacteria and fungi per sample ranging from 240 to 425 and 261 to 463 respectively. These seven forages shared a high number of potentially beneficial taxa, including Bacillus, Pantoea, Candida and Helotiales, but the relative proportion of these taxa was different in each seed. Fungal communities were clustered more distinctively by host genotypes than bacterial. Some bacterial taxa may be involved in the recruitment of genera from the same phylum. Three Pantoea sp. and five Bacillus sp. were isolated from seeds, and all showed positive effects on Medicago sativa germination rate under salt stress, and of these, Bacillus subtilis Es-1 and Pantoea agglomerans Ed-3 performed best, but their influence was affected by the seed's microbiome. Rather than simply promoting host plant growth directly, some taxa may also participate in organizing the assembly of plant microbiomes which will influence seed response to biological factors. This study uses a new, high-throughput sequencing based strategy to identify beneficial strains and analyse the interactions between microorganisms and plants to maximize microbial functions in long-term agricultural practices.

Transcriptome profiles identify the common responsive genes to drought stress in two Elymus species.

Elymus, the largest genus of the Triticeae Dumort, is a forage grass in the Qinghai-Tibetan Plateau, where the climate has gradually become increasingly dry in recent years. To understand the mechanisms of the response to drought stress in Elymus species, we first investigated physiological and biochemical responses to polyethylene glycol (PEG-6000) simulated drought stress in two Elymus species, Elymus nutans and Elymus sibiricus, and found that E. nutans was more tolerant to drought stress than E. sibiricus. De novo transcriptome analysis of these two Elymus species treated with or without 10 % PEG-6000 revealed that a total of 1695 unigenes were commonly regulated by drought treatment in these two Elymus species, with 1614 unigenes up-regulated and 81 unigenes down-regulated. The coexpressed differentially expressed genes (DEGs) were enriched in regulation of transcription and gene expression in the GO database. KEGG pathway analysis indicated plant hormone signaling transduction were mostly enriched in co-expressed DEGs. Furthermore, genes annotated in the plant hormone signaling transduction were screened from co-expressed DEGs, and found that abscisic acid plays the major role in the drought stress tolerance of Elymus. Meanwhile, transcription factors screened from co-expressed DEGs were mainly classified into the ERF subfamily and WRKY, DREB, and HSF family members. Our results provide further reference for studying the response mechanism and culturing highly tolerant grasses of the Elymus species under drought stress.

Mutation of the chloroplast-localized phosphate transporter OsPHT2;1 reduces flavonoid accumulation and UV tolerance in rice.

Phosphorus (P) is an essential macronutrient required for plant development and production. The mechanisms regulating phosphate (Pi) uptake are well established, but the function of chloroplast Pi homeostasis is poorly understood in Oryza sativa (rice). PHT2;1 is one of the transporters/translocators mediating Pi import into chloroplasts. In this study, to gain insight into the role of OsPHT2;1-mediated stroma Pi, we analyzed OsPHT2;1 function in Pi utilization and photoprotection. Our results showed that OsPHT2;1 was induced by Pi starvation and light exposure. Cell-based assays showed that OsPHT2;1 localized to the chloroplast envelope and functioned as a low-affinity Pi transporter. The ospht2;1 had reduced Pi accumulation, plant growth and photosynthetic rates. Metabolite profiling revealed that 52.6% of the decreased metabolites in ospht2;1 plants were flavonoids, which was further confirmed by 40% lower content of total flavonoids compared with the wild type. As a consequence, ospht2;1 plants were more sensitive to UV-B irradiation. Moreover, the content of phenylalanine, the precursor of flavonoids, was also reduced, and was largely associated with the repressed expression of ADT1/MTR1. Furthermore, the ospht2;1 plants showed decreased grain yields at relatively high levels of UV-B irradiance. In summary, OsPHT2;1 functions as a chloroplast-localized low-affinity Pi transporter that mediates UV tolerance and rice yields at different latitudes.

Phosphoproteomic Profiling Reveals the Importance of CK2, MAPKs and CDPKs in Response to Phosphate Starvation in Rice.

Phosphorus is one of the most important macronutrients required for plant growth and development. The importance of phosphorylation modification in regulating phosphate (Pi) homeostasis in plants is emerging. We performed phosphoproteomic profiling to characterize proteins whose degree of phosphorylation is altered in response to Pi starvation in rice root. A subset of 554 proteins, including 546 down-phosphorylated and eight up-phosphorylated proteins, exhibited differential phosphorylation in response to Pi starvation. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis with the differentially phosphorylated proteins indicated that RNA processing, transport, splicing and translation and carbon metabolism played critical roles in response to Pi starvation in rice. Levels of phosphorylation of four mitogen-activated protein kinases (MAPKs), including OsMAPK6, five calcium-dependent protein kinases (CDPKs) and OsCK2ß3 decreased in response to Pi starvation. The decreased phosphorylation level of OsMAPK6 was confirmed by Western blotting. Mutation of OsMAPK6 led to Pi accumulation under Pi-sufficient conditions. Motif analysis indicated that the putative MAPK, casein kinase 2 (CK2) and CDPK substrates represented about 54.4%, 21.5% and 4.7%, respectively, of the proteins exhibiting differential phosphorylation. Based on the motif analysis, 191, 151 and 46 candidate substrates for MAPK, CK2 and CDPK were identified. These results indicate that modification of phosphorylation profiles provides complementary information on Pi-starvation-induced processes, with CK2, MAPK and CDPK protein kinase families playing key roles in these processes in rice.

The role of OsNLA1 in regulating arsenate uptake and tolerance in rice.

Arsenic (As) contamination in agricultural soil can cause phytotoxicity and lead to As accumulation in crops. Rice (Oryza sativa) feeds half of the world's population, but the molecular mechanism of As detoxification is not well understood in rice. In this study, the role of OsNLA1 in arsenate uptake and tolerance in rice was analyzed. OsNLA1 expression was induced in response to As(V) stress. The osnla1 mutant was more sensitive to As(V) stress than those of the wild type (WT). When exposed to As(V), mutation of OsNLA1 resulted in 30% greater As accumulation in roots and shoots of the WT. Although OsPT8 expression was induced after As(V) exposure, the amount of its protein was reduced. Unexpectedly, the osnla1 mutant showed a significant increase in punctate structures of OsPT8-GFP in response to As(V) stress, while the amount of the OsPT8-GFP protein in the osnla1 mutant was greater than in the WT. Combining OsNLA1 mutation with OsPT8 overexpression resulted in As(V) hypersensitivity, As hyperaccumulation, and higher shoot to root ratio of As in rice. These results indicated that OsNLA1 plays an important role in arsenate uptake and tolerance, mainly via regulating the amount of Pi transporters.

Hydrogen Peroxide and Nitric Oxide Crosstalk Mediates Brassinosteroids Induced Cold Stress Tolerance in Medicago truncatula.

Brassinosteroids (BRs) play pivotal roles in modulating plant growth, development, and stress responses. In this study, a Medicago truncatula plant pretreated with brassinolide (BL, the most active BR), enhanced cold stress tolerance by regulating the expression of several cold-related genes and antioxidant enzymes activities. Previous studies reported that hydrogen peroxide (H2O2) and nitric oxide (NO) are involved during environmental stress conditions. However, how these two signaling molecules interact with each other in BRs-induced abiotic stress tolerance remain largely unclear. BL-pretreatment induced, while brassinazole (BRZ, a specific inhibitor of BRs biosynthesis) reduced H2O2 and NO production. Further, application of dimethylthiourea (DMTU, a H2O2 and OH- scavenger) blocked BRs-induced NO production, but BRs-induced H2O2 generation was not sensitive to 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO, a scavenger of NO). Moreover, pretreatment with DMTU and PTIO decreased BL-induced mitochondrial alternative oxidase (AOX) and the photosystem capacity. However, pretreatment with PTIO was found to be more effective than DMTU in reducing BRs-induced increases in Valt, Vt, and MtAOX1 gene expression. Similarly, BRs-induced photosystem II efficiency was found in NO dependent manner than H2O2. Finally, we conclude that H2O2 was involved in NO generation, whereas NO was found to be crucial in BRs-induced AOX capacity, which further contributed to the protection of the photosystem under cold stress conditions in Medicago truncatula.

A Phosphate-Starvation Induced RING-Type E3 Ligase Maintains Phosphate Homeostasis Partially Through OsSPX2 in Rice.

Phosphate (Pi), as the main form of phosphorus that can be absorbed by plants, is one of the most limiting macro-nutrients for plants. However, the mechanism for maintaining Pi homeostasis in rice (Oryza sativa) is still not well understood. We identified a Pi-starvation-induced E3 ligase (OsPIE1) in rice. Using an in vitro self-ubiquitination assay, we determined the E3 ligase activities of OsPIE1. Using GUS staining and GFP detection, we analyzed tissue expression patterns of OsPIE1 and the subcellular localization of its encoded protein. The function of OsPIE1 in Pi homeostasis was analyzed using OsPIE1 overexpressors and ospie1 mutants. OsPIE1 was localized to the nucleus, and expressed in epidermis, exodermis and sclerenchyma layers of primary root. Under Pi-sufficient condition, overexpression of OsPIE1 upregulated the expression of OsPT2, OsPT3, OsPT10 and OsPAP21b, resulting in Pi accumulation and acid phosphatases (APases) induction in roots. OsSPX2 was strongly suppressed in OsPIE1 overexpressors. Further comparative transcriptome analysis, tissue expression patterns and genetic interaction analysis indicated that the enhancing of Pi accumulation and APase activities upon overexpression of OsPIE1 was (at least in part) caused by repression of OsSPX2. These results indicate that OsPIE1 plays an important role in maintaining Pi homeostasis in rice.

N gene enhances resistance to Chilli veinal mottle virus and hypersensitivity to salt stress in tobacco.

Plants use multiple mechanisms to fight against pathogen infection. One of the major mechanisms involves the disease resistance (R) gene, which specifically mediates plant defense. Recent studies have shown that R genes have broad spectrum effects in response to various stresses. N gene is the resistance gene specifically resistant to Tobacco mosaic virus (TMV). However, the role of N gene in abiotic stress and other viral responses remains obscure. In this study, we investigated the mechanisms by which N regulates plant defense responses under Chilli veinal mottle virus (ChiVMV) infection and salt stress. Here, we monitored the physiological and molecular changes of tobacco plants under virus attack. The results showed that when tobaccoNN and tobacconn plants were exposed to ChiVMV, tobaccoNN plants displayed higher susceptibility at five days post infection (dpi), while tobacconn plants exhibited higher susceptibility at 20 dpi. In addition, accumulation of reactive oxygen species (ROS) and expression of HARPIN-INDUCED1(NtHIN1) were higher in tobaccoNN plants than in tobacconn plants at 5 dpi. Interestingly, the pathogenesis-related gene (NtPR1 and NtPR5), the activities of antioxidant enzymes, and the content of salicylic acid (SA) in tobaccoNN plants increased compared with those in tobacconn plants. It was suggested that the N gene induced a hypersensitive response (HR) and enhanced the systemic resistance of plants in response to ChiVMV via the SA-dependent signaling pathway. In addition, the N gene was also induced significantly by salt stress. However, tobaccoNN plants showed hypersensitivity toward increased salt stress, and this hypersensitivity was dependent on abscisic acid and jasmonic acid but not SA. Taken together, our results indicate that the N gene appears to be important in the plant response to ChiVMV infection and salt stress.

α-Expansin EXPA4 Positively Regulates Abiotic Stress Tolerance but Negatively Regulates Pathogen Resistance in Nicotiana tabacum.

Since they function as cell wall-loosening proteins, expansins can affect plant growth, developmental processes and environmental stress responses. Our previous study demonstrated that changes in Nicotiana tabacum α-expansin 4 (EXPA4) expression affect the sensitivity of tobacco to Tobacco mosaic virus [recombinant TMV encoding green fluorescent protein (TMV-GFP)] infection by Agrobacterium-mediated transient expression. In this study, to characterize the function of tobacco EXPA4 further, EXPA4 RNA interfernce (RNAi) mutants and overexpression lines were generated and assayed for their tolerance to abiotic stress and resistance to pathogens. First, the differential phenotypes and histomorphology of transgenic plants with altered EXPA4 expression indicated that EXPA4 is essential for normal tobacco growth and development. By utilizing tobacco EXPA4 mutants with abiotic stress, it was demonstrated that RNAi mutants have increased hypersensitivity to salt and drought stress. In contrast, the overexpression of EXPA4 in tobacco conferred greater tolerance to salt and drought stress, as indicated by less cell damage, higher fresh weight, higher soluble sugar and proline accumulation, and higher expression levels of several stress-responsive genes. In addition, the overexpression lines were more susceptible to the viral pathogen TMV-GFP when compared with the wild type or RNAi mutants. The induction of the antioxidant system, several defense-associated phytohormones and gene expression was down-regulated in overexpression lines but up-regulated in RNAi mutants when compared with the wild type following TMV-GFP infection. In addition, EXPA4 overexpression also accelerated the disease development of Pseudomonas syringae DC3000 on tobacco. Taken together, these results suggested that EXPA4 appears to be important in tobacco growth and responses to abiotic and biotic stress.

Alpha-momorcharin enhances Tobacco mosaic virus resistance in tobaccoNN by manipulating jasmonic acid-salicylic acid crosstalk.

Alpha-momorcharin (α-MMC) is a type-I ribosome inactivating protein (RIP) with a molecular weight of 29 kDa found in plants. This protein has been shown to be effective against a broad range of human viruses and also has anti-tumor activities. However, the mechanism by which α-MMC induces plant defense responses and regulates the N gene to promote resistance to the Tobacco mosaic virus (TMV) is still not clear. By using pharmacological and infection experiments, we found that α-MMC enhances TMV resistance of tobacco plants containing the N gene (tobaccoNN). Our results showed that plants pretreated with 0.5 mg/ml α-MMC could relieve TMV-induced oxidative damage, had enhanced the expression of the N gene and increased biosynthesis of jasmonic acid (JA) and salicylic acid (SA). Moreover, transcription of JA and SA signaling pathway genes were increased, and their expression persisted for a longer period of time in plants pretreated with α-MMC compared with those pretreated with water. Importantly, exogenous application of 1-Aminobenzotriazole (ABT, SA inhibitor) and ibuprofen (JA inhibitor) reduced α-MMC induced plant resistance under viral infection. Thus, our results revealed that α-MMC enhances TMV resistance of tobaccoNN plants by manipulating JA-SA crosstalk.

Nitric oxide as a signaling molecule in brassinosteroid-mediated virus resistance to Cucumber mosaic virus in Arabidopsis thaliana.

Brassinosteroids (BRs) are growth-promoting plant hormones that play a crucial role in biotic stress responses. Here, we found that BR treatment increased nitric oxide (NO) accumulation, and a significant reduction of virus accumulation in Arabidopsis thaliana. However, the plants pre-treated with NO scavenger [2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-1-oxyl-3-oxide (PTIO)] or nitrate reductase (NR) inhibitor (tungstate) hardly had any NO generation and appeared to have the highest viral replication and suffer more damages. Furthermore, the antioxidant system and photosystem parameters were up-regulated in brassinolide (BL)-treated plants but down regulated in PTIO- or tungstate-treated plants, suggesting NO may be involved in BRs-induced virus resistance in Arabidopsis. Further evidence showed that NIA1 pathway was responsible for BR-induced NO accumulation in Arabidopsis. These results indicated that NO participated in the BRs-induced systemic resistance in Arabidopsis. As BL treatment could not increase NO levels in nia1 plants in comparison to nia2 plants. And nia1 mutant exhibited decreased virus resistance relative to Col-0 or nia2 plants after BL treatment. Taken together, our study addressed that NIA1-mediated NO biosynthesis is involved in BRs-mediated virus resistance in A. thaliana.

Tobacco alpha-expansin EXPA4 plays a role in Nicotiana benthamiana defence against Tobacco mosaic virus.

MAIN CONCLUSION: Tobacco EXPA4 plays a role in Nicotiana benthamiana defence against virus attack and affects antioxidative metabolism and phytohormone-mediated immunity responses in tobacco. Expansins are cell wall-loosening proteins known for their endogenous functions in cell wall extensibility during plant growth. The effects of expansins on plant growth, developmental processes and environment stress responses have been well studied. However, the exploration of expansins in plant virus resistance is rarely reported. In the present study, virus-induced gene silencing (VIGS) and Agrobacterium-mediated transient overexpression were conducted to investigate the role of Nicotiana tabacum alpha-expansin 4 (EXPA4) in modulating Tobacco mosaic virus (TMV-GFP) resistance in Nicotiana benthamiana. The results indicated that silencing of EXPA4 reduced the sensitivity of N. benthamiana to TMV-GFP, and EXPA4 overexpression accelerated virus reproduction on tobacco. In addition, our data suggested that the changes of virus accumulation in response to EXPA4 expression levels could further affect the antioxidative metabolism and phytohormone-related pathways in tobacco induced by virus inoculation. EXPA4-silenced plants with TMV-GFP have enhanced antioxidant enzymes activities, which were down-regulated in virus-inoculated 35S:EXPA4 plants. Salicylic acid accumulation and SA-mediated defence genes induced by TMV-GFP were up-regulated in EXPA4-silenced plants, but depressed in 35S:EXPA4 plants. Furthermore, a VIGS approach was used in combination with exogenous phytohormone treatments, suggesting that EXPA4 has different responses to different phytohormones. Taken together, these results suggested that EXPA4 plays a role in tobacco defence against viral pathogens.

Photo-protective mechanisms in reed canary grass to alleviate photo-inhibition of PSII on the Qinghai-Tibet Plateau.

Due to its characteristic of high biomass yield potential, there is considerable interest in cultivating Phalaris arundinacea L. cv. 'chuancaoyin No.3' (reed canary grass) on the Qinghai-Tibet Plateau where there is an abundance of alpine steppe meadow and a potential large market for animal husbandry. In this study, we 1) investigate whether reed canary grass exhibits superior productive capacity to Elymus nutans 'Aba' (E. nutans), ordinary common pasture, during the long warm days of summer at high-altitude; and 2) compare the cold tolerance between reed canary grass and E. nutans, including photosynthesis, photo-inhibition, and photo-protection. The results suggest that reed canary grass exhibits higher photosynthetic capacity compared to E. nutans at latitudes of the cool temperate zone. Meanwhile, cold-induced photo-inhibition and photo-damage at high altitudes in reed canary grass were due to both stomatal and non-stomatal limitation, and the enhancement in photo-respiration, thermal dissipation, and Mehler reaction are important processes to minimize the negative effects of high elevation and a cold environment.

The Roles of Alpha-Momorcharin and Jasmonic Acid in Modulating the Response of Momordica charantia to Cucumber Mosaic Virus.

Alpha-momorcharin (α-MMC) is a type-I ribosome inactivating protein with a molecular weight of 29 kDa that is found in Momordica charantia, and has been shown to be effective against a broad range of human viruses as well as having anti-tumor activities. However, the role of endogenous α-MMC under viral infection and the mechanism of the anti-viral activities of α-MMC in plants are still unknown. To study the effect of α-MMC on plant viral defense and how α-MMC increases plant resistance to virus, the M. charantia-cucumber mosaic virus (CMV) interaction system was investigated. The results showed that the α-MMC level was positively correlated with the resistance of M. charantia to CMV. α-MMC treatment could alleviate photosystem damage and enhance the ratio of glutathione/glutathione disulfide in M. charantia under CMV infection. The relationship of α-MMC and defense related phytohormones, and their roles in plant defense were further investigated. α-MMC treatment led to a significant increase of jasmonic acid (JA) and vice versa, while there was no obvious relevance between salicylic acid and α-MMC. In addition, reactive oxygen species (ROS) were induced in α-MMC-pretreated plants, in a similar way to the ROS burst in JA-pretreated plants. The production of ROS in both ibuprofen (JA inhibitor) and (α-MMC+ibuprofen)-pretreated plants was reduced markedly, leading to a greater susceptibility of M. charantia to CMV. Our results indicate that the anti-viral activities of α-MMC in M. charantia may be accomplished through the JA related signaling pathway.

Medicago truncatula genotypes Jemalong A17 and R108 show contrasting variations under drought stress.

Drought is one of the most significant abiotic stresses that restrict crop productivity. Medicago truncatula is a model legume species with a wide genetic diversity. We compared the differential physiological and molecular changes of two genotypes of M. truncatula (Jemalong A17 and R108) in response to progressive drought stress and rewatering. The MtNCED and MtZEP activation and higher abscisic acid (ABA) content was observed in Jemalong A17 plants under normal conditions. Additionally, a greater increase in ABA content and expression of MtNCED and MtZEP in Jemalong A17 plants than that of R108 plants were observed under drought conditions. A more ABA-sensitive stomatal closure and a slower water loss was found in excised leaves of Jemalong A17 plants. Meanwhile, Jemalong A17 plants alleviated leaf wilting and maintained higher relative water content under drought conditions. Exposed to drought stress, Jemalong A17 plants exhibited milder oxidative damage which has less H2O2 and MDA accumulation, lower electrolyte leakage and higher chlorophyll content and PSII activity. Furthermore, Jemalong A17 plants enhanced expression of stress-upregulated genes under drought conditions. These results suggest that genotypes Jemalong A17 and R108 differed in their response and adaptation to drought stress. Given the relationship between ABA and these physiological responses, the MtNCED and MtZEP activation under normal conditions may play an important role in regulation of greater tolerance of Jemalong A17 plants to drought stress. The activation of MtNCED and MtZEP may lead to the increase of ABA content which may activate expression of drought-stress-regulated genes and cause a series of physiological resistant responses.

Arabidopsis cryptochrome 1 functions in nitrogen regulation of flowering.

The phenomenon of delayed flowering after the application of nitrogen (N) fertilizer has long been known in agriculture, but the detailed molecular basis for this phenomenon is largely unclear. Here we used a modified method of suppression-subtractive hybridization to identify two key factors involved in N-regulated flowering time control in Arabidopsis thaliana, namely ferredoxin-NADP(+)-oxidoreductase and the blue-light receptor cryptochrome 1 (CRY1). The expression of both genes is induced by low N levels, and their loss-of-function mutants are insensitive to altered N concentration. Low-N conditions increase both NADPH/NADP(+) and ATP/AMP ratios, which in turn affect adenosine monophosphate-activated protein kinase (AMPK) activity. Moreover, our results show that the AMPK activity and nuclear localization are rhythmic and inversely correlated with nuclear CRY1 protein abundance. Low-N conditions increase but high-N conditions decrease the expression of several key components of the central oscillator (e.g., CCA1, LHY, and TOC1) and the flowering output genes (e.g., GI and CO). Taken together, our results suggest that N signaling functions as a modulator of nuclear CRY1 protein abundance, as well as the input signal for the central circadian clock to interfere with the normal flowering process.