Background: Mitogen activated protein kinase (MAPK) cascade is an important signaling cascade that operates in stress signal transduction in plants. The biologically active monoterpenoid indole alkaloids (MIA) produced in Catharanthus roseus are known to be induced under several abiotic stress conditions such as wounding, UV-B etc. However involvement of any signaling component in the accumulation of MIAs remains poorly investigated so far. Here we report isolation of a novel abiotic stress inducible Catharanthus roseus MAPK, CrMPK3 that may have role in accumulation of MIAs in response to abiotic stress. Results: CrMPK3 expressed in bacterial system is an active kinase as it showed auto-phosphorylation and phosphorylation of Myelin Basic Protein. CrMPK3 though localized in cytoplasm, moves to nucleus upon wounding. Wounding, UV treatment and MeJA application on C. roseus leaves resulted in the transcript accumulation of CrMPK3 as well as activation of MAPK in C. roseus leaves. Immuno-precipitation followed by immunoblot analysis revealed that wounding, UV treatment and methyl jasmonate (MeJA) activate CrMPK3. Transient over-expression of CrMPK3 in C. roseus leaf tissue showed enhanced expression of key MIA biosynthesis pathway genes and also accumulation of specific MIAs. Conclusion: Results from our study suggest a possible involvement of CrMPK3 in abiotic stress signal transduction towards regulation of transcripts of key MIA biosynthetic pathway genes, regulators and accumulation of major MIAs.
Catharanthus roseus is an important source of pharmaceutically important Monoterpenoid Indole Alkaloids (MIAs). Accumulation of many of the MIAs is induced in response to abiotic stresses such as wound, ultra violet (UV) irradiations, etc. Recently, we have demonstrated a possible role of CrMPK3, a C. roseus mitogen-activated protein kinase in stress-induced accumulation of a few MIAs. Here, we extend our findings using Saccharomyces cerevisiae to investigate the role of CrMPK3 in giving tolerance to abiotic stresses. Yeast cells transformed with CrMPK3 was found to show enhanced tolerance to UV and heat stress. Comparison of CrMPK3 and SLT2, a MAPK from yeast shows high-sequence identity particularly at conserved domains. Additionally, heat stress is also shown to activate a 43 kDa MAP kinase, possibly CrMPK3 in C. roseus leaves. These findings indicate the role of CrMPK3 in stress-induced MIA accumulation as well as in stress tolerance.
Grain yield of wheat is primarily determined by both grain number and grain weight, which often influence each other in response to environmental stimuli. Some of the genotypes are capable of maintaining high single grain weight (SGW) across the environments. Understanding mechanisms and factors associated with the superiority of such genotypes over others is necessary to enhance productivity of wheat. Experiments were conducted to elucidate the physiological basis of high SGW of LOK-1, a wheat cultivar grown in dry and hot environments in the central and peninsular zones of India. SGW of LOK-1 was least affected by removal of spikelets indicating little competition between the grains within the spike for assimilates. Reduction in SGW due to defoliation was less and the contribution of stem reserves to the grain development was high in LOK-1 relative to other cultivars. It seems that high level of expression of genes such as expansin (TaExpA6) contributes to the high SGW of LOK-1. Source was not a limiting factor for grain growth of LOK-1 in contrast to other cultivars, whereas sink appeared to be a limiting factor in recently released/identified cultivars. Differences in the amounts of water soluble stem carbohydrate reserves translocated to grain could be one of the factors contributing to higher grain weight in LOK-1. High level expression of TaExpA6, one of the genes contributing to the elongation of endosperm, seems to be crucial for grain growth in wheat.