Nabil I. Elsheery

Frequently asked questions about in vivo chlorophyll fluorescence: practical issues

The aim of this educational review is to provide practical information on the hardware, methodology, and the hands on application of chlorophyll (Chl) a fluorescence technology. We present the paper in a question and answer format like frequently asked questions. Although nearly all information on the application of Chl a fluorescence can be found in the literature, it is not always easily accessible. This paper is primarily aimed at scientists who have some experience with the application of Chl a fluorescence but are still in the process of discovering what it all means and how it can be used. Topics discussed are (among other things) the kind of information that can be obtained using different fluorescence techniques, the interpretation of Chl a fluorescence signals, specific applications of these techniques, and practical advice on different subjects, such as on the length of dark adaptation before measurement of the Chl a fluorescence transient. The paper also provides the physiological background for some of the applied procedures. It also serves as a source of reference for experienced scientists.

Identification and expression analysis of two small heat shock protein cDNAs from developing seeds of biodiesel feedstock plant Jatropha curcas

Plant small heat shock proteins (sHSPs) are known to be important for environmental stress tolerance and involved in various developmental processes. In this study, two full-length cDNAs encoding sHSPs, designated JcHSP-1 and JcHSP-2, were identified and characterized from developing seeds of a promising biodiesel feedstock plant Jatropha curcas by expressed sequence tag (EST) sequencing of embryo cDNA libraries and rapid amplification of cDNA ends (RACE). JcHSP-1 and JcHSP-2 contained open-reading frames encoding sHSPs of 219 and 157 amino acids, with predicted molecular weights of 24.42kDa and 18.02kDa, respectively. Sequence alignment indicated that both JcHSP-1 and JcHSP-2 shared high similarity with other plant sHSPs. Real-time quantitative RT-PCR analysis showed that the transcriptional level of both JcHSP-1 and JcHSP-2 increased along with natural dehydration process during seed development. A sharp increase of JcHSP-2 transcripts occurred in response to water content dropping from 42% in mature seeds to 12% in dry seeds. Western blot analysis revealed that the accumulation profile of two cross-reacting proteins, whose molecular weight corresponding to the calculated size of JcHSP-1 and JcHSP-2, respectively, was well consistent with the mRNA expression pattern of JcHSP-1 and JcHSP-2 in jatropha seeds during maturation and natural dehydration. These results indicated that both JcHSPs might play an important role in cell protection and seed development during maturation of J. curcas seeds.

Regulation and physiological role of silicon in alleviating drought stress of mango

Improvement of drought stress of mango plants requires intensive research that focuses on physiological processes. In three successive seasons (2014, 2015and 2016) field experiments with four different strains of mango were subjected to two water regimes. The growth and physiological parameters of possible relevance for drought stress tolerances in mango were investigated. Yield and its components were also evaluated. The data showed that all growth and physiological parameters were increased under K2SiO3 (Si) supplement and were followed by the interaction treatment (Si treatment and its combination with drought stress) compared to that of the controlled condition. Drought stress decreased the concentration of auxins (IAA), gibberellins (GA) and cytokinins (CK) in the three mango cultivars leaves, whereas, it increased the concentration of abscisic acid (ABA). On the contrary, IAA, GA, and CK (promoters) endogenous levels were improved by supplementing Si, in contrary ABA was decreased. Drought stress increased the activity of peroxidase (POX), catalase (CAT), and superoxide dismutase (SOD) in the leaves of all mango cultivars grown during three experimental seasons. However, Si supplementation reduced the levels of all these antioxidative enzymes, especially the concentration of SOD when compared to that of control leaves. Fruit quality was improved in three successive seasons when Si was applied. Our results clearly show that the increment in drought tolerance was associated with an increase in antioxidative enzyme activity, allowing mango plants to cope better with drought stress. Si possesses an efficient system for scavenging reactive oxygen species, which protects the plant against destructive oxidative reactions, thereby improving the ability of the mango trees to withstand environmental stress in arid regions.

Identification and differential expression of two dehydrin cDNAs during maturation of Jatropha curcas seeds

Plant dehydrin proteins (DHNs) are known to be important for environmental stress tolerance and are involved in various developmental processes. Two full-length cDNAs JcDHN-1 and JcDHN-2 encoding two dehydrins from Jatropha curcas seeds were identified and characterized. JcDHN-1 is 764 bp long and contains an open reading frame of 528 bp. The deduced JcDHN-1 protein has 175 a.a. residues that form a 19.3-kDa polypeptide with a predicted isoelectric point (pI) of 6.41. JcDHN-2 is 855 bp long and contains an open reading frame of 441 bp. The deduced JcDHN-2 protein has 156 a.a. residues that form a 17.1-kDa polypeptide with a predicted pI of 7.09. JcDHN-1 is classified as type Y3SK2 and JcDHN-2 is classified as type Y2SK2 according to the YSK shorthand for structural classification of dehydrins. Homology analysis indicates that both JcDHN-1 and JcDHN-2 share identity with DHNs of other plants. Analysis of the conserved domain revealed that JcDHN-2 has glycoside hydrolase GH20 super-family activity. Quantitative real time PCR analysis for JcDHN-1 and JcDHN-2 expression during seed development showed increasing gene expression of both their transcript levels along with the natural dehydration process during seed development. A sharp increase in JcDHN-2 transcript level occurred in response to water content dropping from 42% in mature seeds to 12% in dry seeds. These results indicate that both JcDHNs have the potential to play a role in cell protection during dehydration occurring naturally during jatropha orthodox seed development.

Dehydroascorbate reductase and glutathione reductase play an important role in scavenging hydrogen peroxide during natural and artificial dehydration of Jatropha curcas seeds

Changes in H2O2 and the main antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and glutathione reductase (GR), in endospermic and embryonic tissues were studied in developing and artificially dried Jatropha curcas seeds. Immature seeds were desiccation-tolerant at 80 days after flowering, as they were able to germinate fully after artificial drying on silica gel had reduced their water content to 10–12% of fresh weight. In both endospermic and embryonic tissues, H2O2 level and, consequently, lipid peroxide content, decreased during seed development as well as after artificial dehydration of developing seeds. All examined antioxidant enzymes except DHAR showed a decrease in total activity in mature stages as compared with early stages. Expression analysis of SOD genes revealed that the decrease in total SOD activities was related to the decrease in Cu/Zn-SOD expression, while the continuous activity of SOD during maturation was related to an increase in Mn-SOD expression. Artificial drying resulted in increased SOD and DHAR activity, irrespective of the developmental stage. Our results revealed weak participation of CAT and APX in H2O2 scavenging, as well as no significant alterations in GR activities either during maturation or after artificial drying. Changes in SOD and GR isoenzyme patterns occurred during maturation-related drying, but not after artificial drying. These results highlight the role of ascorbate-glutathione cycle enzymes (DHAR and GR) in H2O2 scavenging during maturation or after artificial drying of developing J. curcas seeds.