Molecular Characterization of Mg-Chelatase CHLI Subunit in Pea (Pisum sativum L.).

As a rate-limiting enzyme for chlorophyll biosynthesis, Mg-chelatase is a promising target for improving photosynthetic efficiency. It consists of CHLH, CHLD, and CHLI subunits. In pea (Pisum sativum L.), two putative CHLI genes (PsCHLI1 and PsCHLI2) were revealed recently by the whole genome sequencing, but their molecular features are not fully characterized. In this study, PsCHLI1 and PsCHLI2 cDNAs were identified by PCR-based cloning and sequencing. Phylogenetic analysis showed that PsCHLIs were derived from an ancient duplication in legumes. Both PsCHLIs were more highly expressed in leaves than in other organs and downregulated by abscisic acid and heat treatments, while PsCHLI1 was more highly expressed than PsCHLI2. PsCHLI1 and PsCHLI2 encode 422- and 417-amino acid proteins, respectively, which shared 82% amino acid identity and were located in chloroplasts. Plants with a silenced PsCHLI1 closely resembled PsCHLI1 and PsCHLI2 double-silenced plants, as both exhibited yellow leaves with barely detectable Mg-chelatase activity and chlorophyll content. Furthermore, plants with a silenced PsCHLI2 showed no obvious phenotype. In addition, the N-terminal fragment of PsCHLI1 (PsCHLI1N, Val63-Cys191) and the middle fragment of PsCHLI1 (PsCHLI1M, Gly192-Ser336) mediated the formation of homodimers and the interaction with CHLD, respectively, while active PsCHLI1 was only achieved by combining PsCHLI1N, PsCHLI1M, and the C-terminal fragment of PsCHLI1 (Ser337-Ser422). Taken together, PsCHLI1 is the key CHLI subunit, and its peptide fragments are essential for maintaining Mg-chelatase activity, which can be used to improve photosynthetic efficiency by manipulating Mg-chelatase in pea.

Red Light-Induced Systemic Resistance Against Root-Knot Nematode Is Mediated by a Coordinated Regulation of Salicylic Acid, Jasmonic Acid and Redox Signaling in Watermelon.

Red light (RL) can stimulate plant defense against foliar diseases; however, its role in activation of systemic defense against root diseases remains unclear. Here, the effect of RL on root knot nematode Meloidogyne incognita (RKN) infestation was investigated in watermelon plants (Citrullus lanatus L.). Plants were exposed to 200 µmol m-2 s-1 photosynthetic photon flux density RL at the canopy level for 21 days using light-emitting photodiodes. The results showed that RL significantly suppressed gall formation and nematode development, which was closely associated with the RL-induced attenuation of oxidative stress in roots. Gene expression analysis showed that RL caused a transient upregulation of PR1 and WRKY70 transcripts at 7 days post inoculation in RKN-infected plants. Further investigation revealed that RL-induced systemic defense against RKN was attributed to increased jasmonic acid (JA) and salicylic acid (SA) content, and transcript levels of their biosynthetic genes in roots. Interestingly, while malondialdehyde content decreased, H2O2 accumulation increased in RL-treated RKN-plants, indicating a potential signaling role of H2O2 in mediating RL-induced systemic defense. Furthermore, analysis of enzymatic and non-enzymatic antidoxidants revealed that RL-induced enhanced defense agaist RKN was also attributed to increased activities of antioxidant enzymes as well as redox homeostasis. Taken together, these findings suggest that RL could enhance systemic resistance against RKN, which is mediated by a coordinated regulation of JA- and SA-dependent signaling, antioxidants, and redox homeostasis in watermelon plants.

Crosstalk among Jasmonate, Salicylate and Ethylene Signaling Pathways in Plant Disease and Immune Responses.

Phytohormone crosstalk is crucial for plant defenses against pathogens and insects in which salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) play key roles. These low molecular mass signals critically trigger and modulate plant resistance against biotrophic as well as necrotrophic pathogens through a complex signaling network that even involves participation of other hormones. Crosstalk among SA, JA and ET is mediated by different molecular players, considered as integral part of these crosscommunicating signal transduction pathways. Recent progress has revealed that the positive versus negative interactions among those pathways ultimately enable a plant to fine-tune its defense against specific aggressors. On the other hand, pathogens have evolved strategies to manipulate the signaling network to their favour in order to intensify virulence on host plant. Here we review recent advances and current knowledge on the role of classical primary defense hormones SA, JA and ET as well as their synergistic and antagonistic interaction in plant disease and immune responses. Crosstalk with other hormones such as abscisic acid, auxin, brassinosteroids, cytokinins and melatonin is also discussed mainly in plant disease resistance. In addition to our keen focus on hormonal crosstalk, this review also highlights potential implication of positive and negative regulatory interactions for developing an efficient disease management strategy through manipulation of hormone signaling in plant.

RNA-seq analysis reveals the role of red light in resistance against Pseudomonas syringae pv. tomato DC3000 in tomato plants.

BACKGROUND: Plants attenuate their responses to a variety of bacterial and fungal pathogens, leading to higher incidences of pathogen infection at night. However, little is known about the molecular mechanism responsible for the light-induced defence response; transcriptome data would likely facilitate the elucidation of this mechanism. RESULTS: In this study, we observed diurnal changes in tomato resistance to Pseudomonas syringae pv. tomato DC3000 (Pto DC3000), with the greatest susceptibility before midnight. Nightly light treatment, particularly red light treatment, significantly enhanced the resistance; this effect was correlated with increased salicylic acid (SA) accumulation and defence-related gene transcription. RNA-seq analysis revealed that red light induced a set of circadian rhythm-related genes involved in the phytochrome and SA-regulated resistance response. The biosynthesis and signalling pathways of multiple plant hormones (auxin, SA, jasmonate, and ethylene) were co-ordinately regulated following Pto DC3000 infection and red light, and the SA pathway was most significantly affected by red light and Pto DC3000 infection. This result indicates that SA-mediated signalling pathways are involved in red light-induced resistance to pathogens. Importantly, silencing of nonexpressor of pathogensis-related genes 1 (NPR1) partially compromised red light-induced resistance against Pto DC3000. Furthermore, sets of genes involved in redox homeostasis (respiratory burst oxidase homologue, RBOH; glutathione S-transferases, GSTs; glycosyltransferase, GTs), calcium (calmodulin, CAM; calmodulin-binding protein, CBP), and defence (polyphenol oxidase, PPO; nudix hydrolase1, NUDX1) as well as transcription factors (WRKY18, WRKY53, WRKY60, WRKY70) and cellulose synthase were differentially induced at the transcriptional level by red light in response to pathogen challenge. CONCLUSIONS: Taken together, our results suggest that there is a diurnal change in susceptibility to Pto DC3000 with greatest susceptibility in the evening. The red light induced-resistance to Pto DC3000 at night is associated with enhancement of the SA pathway, cellulose synthase, and reduced redox homeostasis.

[The more efficacious acupoints of Zusanli and Sanyinjiao than that of non-acupoints on bone mass in osteopenic ovariectomized rats].

OBJECTIVE: To clarify whether the acupoints of Zusanli (ST36) and Sanyinjiao (SP6) have specific actions other than non-acupoints to bone. METHODS: Forty Sprague-Dawley female rats were divided into five groups: Sham operated (sham) group; Ovariectomized (OVX, model) group; non-acupuncture group; OVX, needling on Zusanli and Sanyinjiao (Acp-A) group; OVX, needling on the reverse sides of Zusanli and Sanyinjiao (Acp-B) group; OVX, periostineal stimulation on the same height as points of Zusanli and Sanyinjiao (Acp-C) group. The experiment was continued for 23 weeks and then all animals were sacrificed. RESULTS: OVX had a significantly higher body weight and lower bone mineral density (BMD) on the lumbar vertebrae, total femora and tibiae than sham rats, however, Acp-A showed a higher BMD compared with the other OVX groups. On the other hand, bone weights, bone strength and bone morphometry such as trabecular volume, trabecular separation, labeled width and bone formation rate also showed the same improvements in Acp-A as compared to the other OVX rats. CONCLUSION: The stimulation on Zusanli and Sanyinjiao specifically prevented the development of osteopenic rats compared with non-acupoints.