Fine-tuning of DREB1D expression in stomatal guard cells under water deficit is mediated differentially by promoter haplotypes from sensitive and tolerant coffee genotypes.

The aim of this work was to study the regulation of coffee DREB-like genes in leaves of C. arabica subjected to cold, heat, low relative humidity, exogenous abscisic acid and high light stress, as well as in leaves and roots of drought-tolerant and droughtsusceptible clones of Coffea canephora subjected to water limitation. In C. arabica, CaERF017 was the most expressed gene under low temperatures and relative humidity, while low humidity and high temperatures up-regulated the expression of CaERF053 and CaERF014, respectively. Under water limitation, CcDREB1B, CcRAP2.4, CcERF027, CcDREB1D and CcTINY were the most expressed genes mainly in leaves of drought-tolerant C. canephora. On the other hand, expression of the CcERF016, CcRAP2.4 and CcDREB2F genes was highly up-regulated under water limitation in the roots of drought-susceptible C. canephora clone 22. We previously reported fine-tuned regulation of CcDREB1D promoter haplotypes (HP15, HP16 and HP17) in transgenic C. arabica subjected to low humidity. Here, we investigated the regulation of these haplotypes under high light, cold, heat, and abscisic acid (ABA) stress. In apical buds and leaf guard cells, GUS-stained percentages were higher in pHP16L-transformed plants subjected to low humidity, high light and ABA stress than in pHP17L-and pHP15L-transformed plants. We also reported up-regulated expression of the endogenous CaDREB1D gene for both the cold and low humidity in leaves of pHP16L-transformed C. arabica suggesting a key role of this gene in controlling the responses of coffee plants to abiotic stress probably through an ABA-dependent pathway.Keywords Abiotic stress . Abscisic acid -coffee -DREB -gene expression -promoter Key message: DREB-like genes are differentially expressed in drought-tolerant and susceptible clones of C. canephora subjected to different abiotic stress. In C. arabica, both cold, low humidity and ABA up-regulate CaDREB1D gene expression.

The employment of biotechnology-based approaches such as somatic embryogenesis has been applied to several plants including Coffea sp. Despite the economic importance of this genus, few information about the role of key regulatory genes in somatic embryogenesis in coffee is available. This work provides information about ABI3 and VAL2 genes performance by RT-qPCR during indirect somatic embryogenesis of Coffea arabica. To achieve this, bioinformatics analysis was performed to identify the genes of the B3 superfamily in the coffee genome. The cell suspensions lines presented similar histological and regeneration patterns, yielding of up to 6.6 embryos per 1 mg of embryogenic aggregates at 7 months. We have identified possible orthologs for VAL2 (Cc06g00410) and ABI3 (Cc01g17380) as well as the other members belonging to superfamily B3. The CaABI3 expression was higher in dedifferentiated competent cells for somatic embryogenesis as compared with non-embryogenic calli. Whereas the expression of VAL2 gene is more active in cotyledonary embryos and plantlets, showing its clear performance in the embryogenesis late stages. The present study suggests that CaABI3 gene could be potentially used as a biomarker for embryogenic process improvement. The good plantlets development obtained from the protocol used may be a reflection of the high expression of CaVAL2 in cotyledonary embryos and plantlets. Key message The activity of CaABI3 is correlated to embryogenic potential with highly expressed in embryogenic masses and expression of the VAL2 gene is increased at the end of the embryogenic process.

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The aim of this study was to identify the correlation between photochemical efficiency and candidate genes expression to elucidate the drought tolerance mechanisms in coffee progenies (Icatu Vermelho IAC 3851-2 × Catimor UFV 1602-215) previously identified as tolerant in field conditions. Four progenies (2, 5, 12 and 15) were evaluated under water-deficit conditions (water deficit imposed 8 months after transplanting seedlings to the pots) and under irrigated system. Evaluations of physiological parameters and expression of candidate genes for drought tolerance were performed. Progeny 5 showed capacity to maintain water potential, which contributed to lower qP variation between irrigated and deficit conditions. However, the increases of qN and NPQ in response to stress indicate that this progeny is photochemically responsive to small variations of Ψam protecting the photosystem and maintaining qP. Data obtained for progeny 12 indicated a lower water status maintenance capacity, but with increased qN and NPQ providing maintenance of the ɸPSII and ETR parameters. A PCA analysis revealed that the genes coding regulatory proteins, ABA-synthesis, cellular protectors, isoforms of ascorbate peroxidase clearly displayed a major response to drought stress and discriminated the progenies 5 and 12 which showed a better photochemical response. The genes CaMYB1, CaERF017, CaEDR2, CaNCED, CaAPX1, CaAPX5, CaGolS3, CaDHN1 and CaPYL8a were up-regulated in the arabica coffee progenies with greater photochemical efficiency under deficit and therefore contributing to efficiency of the photosynthesis in drought tolerant progenies.

The present study aimed to evaluate the quality of cell suspension of Coffea arabica cv. Catiguá at four different ages by morphological and gene expression analysis. For this purpose, cell suspension samples were collected at 30, 45, 60 and 75 days. Morphological analysis revealed the presence of embryogenic regions in all samples, and after 60 days, non-embryogenic regions were also identified. The genic analyses revealed that as the cells were transferred to a new medium a change in both their physical and chemical conditions was noticed that caused stress. The decrease in SERK and BBM genes expression after 75th day may also be due to the non-embryogenic regions, characterized by large, elongated and vacuolated cells that were observed in the periphery of embryogenic regions, starting from 60 days of culture.

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Biotechnological techniques have been extensively studied to provide practical results for coffee improvement. Among these techniques, the present study deals with the somatic embryogenesis for crop improvement. Aims: To describe and correlate the morphology of Coffea arabica cell suspensions with their growth curve through electron microscopy and photonic analysis. Methodology: During 28 days of the culture period, samples of cell culture were collected at four days interval and the growth curve of the cell suspensions was performed by calculating the growth and the growth rate of the same. Cell suspension samples were used to perform the histological and ultrastructural analyses.