Desiccation tolerance in bryophytes： the rehydration proteomes of Bryum argenteum provide insights into the resuscitation mechanism

Bryum argenteum Hedw. is a desiccation tolerant bryophyte and belongs to one of the most important components of the biological soil crusts （BSCs） found in the deserts of Central Asia. Limited information is available on rehydration-responsive proteins in desiccation tolerant plants. As a complement to our previous research analyzing the rehydration transcriptome, we present a parallel quantitative proteomic effort to study rehydration-responsive proteins. Bryophyte gametophores were desiccated （Dry） and rehydrated for 2 h （R2） and 24 h （R24）. Proteins from Dry, R2 and R24 gametophores were labeled by isobaric tags for relative and absolute quantitation （iTRAQ） to determine the relative abundance of rehydration-responsive proteins. A total of 5503 non-redundant protein sequences were identified and 4772 （86.7%） protein sequences were annotated using Gene Ontology （GO） terms and Pfam classifications. Upon rehydration 239 proteins were elevated and 461 proteins were reduced as compared to the desiccated protein sample. Differentially up-regulated proteins were classified into a number of categories including reactive oxygen species scavenging enzymes, detoxifying enzymes, Late Embryogenesis Abundant （LEA） proteins, heat shock proteins, proteasome components and proteases, and photosynthesis and translation related proteins. Furthermore, the results of the correlation between transcriptome and proteome revealed the discordant changes in the expression between protein and mRNA.

Rates of oxygen consumption and tolerance of hypoxia and desiccation in Chinese black sleeper(Bostrichthys sinensis)and mudskipper(Boleophthalmus pectinirostris)embryos

The rates of oxygen consumption, tolerance of hypoxia and desiccation of the Chinese black sleeper （ Bostrichthys sinensis） and mudskipper （Boleophthalmus pectinirostris） embryos were investigated. The pattern of oxygen consumption of the Chinese black sleeper embryos was similar to that of the mudskipper ones. The lowest rates of oxygen consumption [ （ 1.65 ±0. 66） nmol/ （ind. ·h） ] of the Chinese black sleeper embryos 16 h after fertilization and the lowest rates of oxygen consumption [ （0.79± 0.08 ）nmolf（ ind. · h） ] of the mudskipper embryos 6 h after fertilization were recorded, respectively. Then the rates of oxygen consumption of these two species embryos increased gradually until hatching [ （8.26 ± 1.70 ） nmolf（ ind.· h） in the Chinese black sleeper, （2.69 ± 0.23 ）nmolf（ ind. · h） in mudskipper]. After exposure to hypoxia water （0.16 mg/dm^3）, bradycardia of the embryos occurred in both the Chinese black sleeper and the mudskipper. However, the Chinese black sleeper embryos survived approximately 45 min longer than the mudskipper ones. After exposure to desiccation at a relative humidity of 58%, bradycardia of the embryos was observed in both the Chinese black sleeper and the mudskipper, and the Chinese black sleeper embryos lived approximately 9 min longer than the mudskipper ones.

Electron paramagnetic resonance study of amphiphiles partitioning behavior in desiccation-tolerant moss during dehydration

Desiccation tolerance is a crucial characteristic for desert moss surviving in arid regions. Desiccation procedure always induces amphiphiles transferring from the polar cytoplasm into lipid bodies. The behavior of amphiphiles transferring can contribute to the enhancement of desiccation tolerance and the reduction of plasma membrane integrity simultaneously. The effects of amphiphiles partitioning into the lipid phase during water loss has been studied for pollen and seeds using electron paramagnetic resonance （EPR） spectroscopy. However, desiccation-tolerant high plants occur among mosses, several angiosperms and higher plants seeds or pollens. They have different strategies for survival in dehydration and rehydration. A desiccation-tolerant moss Tortula desertorurn was used to investigate the behaviors of amphiphilic molecules during drying by spin label technology. There are small amount of amphiphilic probes partitioning into membrane during moss leaves dehydration, comparing with that in higher plants. Cytoplasm viscosity changed from 1.14 into glass state only dehydration less than 60 min. Moss leaves lost plasma membrane integrity slightly,from 0.115 to 0.237, occurred simultaneously with amphiphiles partition. The results showed the more advantages of mosses than higher plants in adapting fast dehydration. We propose that EPR spin label is feasible for studying the amphiphiles partitioning mechanisms in membrane protection and damage for desiccation-tolerant mosses.

Downregulation of NADPH-cytochrome P450 reductase via RNA interference increases the susceptibility of Acyrthosiphon pisum to desiccation and insecticides

Nicotinamide adenine dinucleotide phosphate(NADPH)-cytochrome P450 reductase(CPR)is involved in the metabolism of endogenous and exogenous substances,and detoxification of insecticides.RNA interference(RNAi)of CPR in certain insects causes developmental defects and enhanced susceptibility to insecticides.However,the CPR of Acyrthosiphon pisum has not been characterized,and its function is still not understood.In this study,we investigated the biochemical functions of A.pisum CPR(ApCPR).ApCPR was found to be transcribed in all developmental stages and was abundant in the embryo stage,and in the gut,head,and abdominal cuticle.After optimizing the dose and silencing duration of RNAi for downregulating ApCPR,we found that ApCPR suppression resulted in a significant decrease in the production of cuticular and internal hydrocarbon contents,and of cuticular waxy coatings.Deficiency in cuticular hydrocarbons(CHCs)decreased the survival rate of A.pisum under desiccation stress and increased its susceptibility to contact insecticides.Moreover,desiccation stress induced a significant increase in ApCPR mRNA levels.We further confirmed that ApCPR participates in CHC production.These results indicate that ApCPR modulates CHC production,desiccation tolerance,and insecticide susceptibility in A.pisum,and presents a novel target for pest control.

Dormancy Breaking and Storage Behavior of Garcinia cowa Roxb. （Guttiferae） Seeds： Implications for Ecological Function and Germplasm Conservation

Abstract: The dormancy breaking and storage behavior of Garcinia cowa Roxb. seeds were investigated. The seeds of G. cowa had 8–11 months dormancy in their natural habitat. Seeds were matured and dispersed at the end of the rainy season (mid-late August to late September) and were scatter-hoarded by rodents as food for winter after the seeds had fallen to the ground. Seedlings often emerged in the forest during the rainy season (May to August) the following year. Intact seeds of G. cowa failed to germinate after being sown at 30 °C for 120 d and the mean germination time (MGT) of seeds cultured in a shade (50% sunlight) nursery was 252 d. The most effective method of breaking dormancy was to remove the seed coat totally, which reduced the MGT to 13 d at 30 °C. Germination was also promoted by partial removal of the seed coat (excising the hilum and exposing the radicle) and chemical scarification (immersion in 1% H2O2 for 1 d). Unscarified seeds take up water rapidly in the first 96 h, but water was absorbed by the outside seed coat, without penetrating through it. The moisture content (MC) of G. cowa seeds was high (50% in fresh weight) at shedding. The seeds could tolerate desiccation to some extent, until the MC reached approximately 40%; below that, the viability decreases rapidly and all seeds died at approximately 17% of MC. Seed viability decreased rapidly when seeds were chilled at 4 °C; germination was 2% after storage for 1 week. Even stored at 10 °C, seeds began to be damaged after 4 weeks. Seed storage for 1 yr revealed that in both dry (relative humidity (35 ± 5)%) and moist (wet sand) storage conditions, seed viability declined, but germination percentages for seeds stored under moist conditions are better than for seed stored under dry conditions. Because of their low tolerance to desiccation, marked chilling sensitivity and relatively short lifespan, G. cowa seeds should be classified into the tropical recalcitrant category. The ecological implications of dormant recalcitrant seeds and cues on storing recalcitrant seeds were discussed.

Melanin synthesis genes BgTH and BgDdc affect body color and cuticle permeability in Blattella germanica

Melanin is involved in cuticle pigmentation and sclerotization of insects,which is critical for maintaining structural integrity and functional completeness of insect cuti-cle.The 2 key enzymes of tyrosine hydroxylase(TH)and dopa decarboxylase(DDC)predicted in melanin biosynthesis are usually conserved in insects.However,it is unclear whether their function is related to epidermal permeability.In this study,we identified and cloned the gene sequences of BgTH and BgDdc from Blattella germanica,and revealed that they both showed a high expression at the molting,and BgTH was abundant in the head and integument while BgDdc was expressed highest in the fat body.Using RNA in-terference(RNAi),we found that knockdown of BgTH caused molting obstacles in some cockroaches,with the survivors showing pale color and softer integuments,while knock-down of BgDdc was viable and generated an abnormal light brown body color.Desiccation assay showed that the dsBgTH-injected adults died earlier than control groups under a dry atmosphere,but dsBgDdc-injected cockroaches did not.In contrast,when dsRNA-treated cockroaches were reared under a high humidity condition,almost no cockroaches died in all treatments.Furthermore,with eosin Y staining assay,we found that BgTH-RNAi resulted in a higher cuticular permeability,and BgDdc-RNAi also caused slight dye pen-etration.These results demonstrate that BgTH and BgDdc function in body pigmentation and affect the waterproofing ability of the cuticle,and the reduction of cuticular perme-ability may be achieved through cuticle melanization.