We and others found two polymorphic LRRK2 (leucine-rich repeat kinase 2) variants (rs34778348:G>A; p.G2385R and rs33949390:G>C; p.R1628P) associated with Parkinson disease (PD) among Chinese patients, but the common worldwide rs34637584:G>A; p.G2019S mutation, was absent. Focusing exclusively on Han Chinese, we first sequenced the coding regions in young onset and familial PD patients and identified 59 variants. We then examined these variants in 250 patients and 250 control subjects. Among the 17 polymorphic variants, five demonstrated different frequency in cases versus controls and were considered in a larger sample of 1,363 patients and 1,251 control subjects. The relative risk of an individual with both p.G2385R and p.R1628P is about 1.9, and this is reduced to 1.5-1.6 if the individual also carries rs7133914:G>C; p.R1398H or rs7308720:C>A: p.N551K. The risk of a carrier with p.R1628P is largely negated if the individual also carries p.R1398H or p.N551K. In dopaminergic neuronal lines, p.R1398H had significantly lower kinase activity, whereas p.G2385R and p.R1628P showed higher kinase activity than wild type. We provided the first evidence that multiple LRRK2 variants exert an individual effect and together modulate the risk of PD among Chinese.
Effects of four host plants, tobacco, Chinese cabbage, cowpea and sweet potato, on larval and pupal development and survival, and longevity and fecundity of adults of Spodoptera litura (F) (Lepidoptera: Noctuidae), were studied under laboratory conditions (26° C, 60–80% RH), as was the utilization of the four host plants and adaptation on tobacco. All of the biological parameters included in the study were affected by the host plants. In a choice test, S. litura females oviposited most on Chinese cabbage, least on tobacco, and intermediate on cowpea and sweet potato. S. litura larvae developed differently on the four host plants, from shortest to longest in the following order: Chinese cabbage, cowpea, sweet potato, and tobacco. Pupal development was shorter on cowpea than on the other three host plants, and males generally developed longer than females. More females than males were found among emerged adults, and male adults lived 1–2 d longer than females. Larvae survived best on cowpea (81.6%), followed by Chinese cabbage (75.5%), then sweet potato (66.1%), and worst on tobacco (49.2%). Pupal survival rates were relatively high (91.4 – 95.9%) in all four host plant treatments, although that on sweet potato was lower than those on the other three host plants. Pupal weights on tobacco and sweet potato were similar, but both were lower than those on Chinese cabbage and cowpea. Generally, male pupae weighed less than female pupae. Numbers of eggs oviposited by female S. litura were highest on sweet potato, followed by those on cowpea, Chinese cabbage, and lowest on tobacco. Relative food consumption rate was highest on sweet potato, followed by that on cowpea, Chinese cabbage, and lowest on tobacco. In contrast, S. litura larvae that fed on tobacco had higher efficiency of conversion of digested food, highest efficiency of conversion of ingested food, and lowest approximate digestibility as compared with larvae that fed on other host plants. The potential causes for S. litura outbreaks on tobacco are discussed.
As an important oilseed worldwide, Camelina sativa is being increasingly explored for its use in production of food, feed, biofuel and industrial chemicals. However, detailed mechanisms of camelina oil biosynthesis and accumulation, particularly in vegetative tissues, are understood to a very small extent. Here, we present genome-wide identification, cloning and functional analysis of phospholipid diacylglycerol acyltransferase (PDAT) in C. sativa, which catalyses the final acylation step in triacylglycerol (TAG) biosynthesis by transferring a fatty acyl moiety from a phospholipid to diacylglycerol (DAG). We identified five genes (namely CsPDAT1-A, B, and C and CsPDAT2-A and B) encoding PDATs from the camelina genome. CsPDAT1-A is mainly expressed in seeds, whereas CsPDAT1-C preferentially accumulates in flower and leaf tissues. High expression of CsPDAT2-A and CsPDAT2-B was detected in stem and root tissues, respectively. Cold stress induced upregulation of CsPDAT1-A and CsPDAT1-C expression by 3.5- and 2.5-fold, respectively, compared to the control. Salt stress led to an increase in CsPDAT2-B transcripts by 5.1-fold. Drought treatment resulted in an enhancement of CsPDAT2-A mRNAs by twofold and a reduction of CsPDAT2-B expression. Osmotic stress upregulated the expression of CsPDAT1-C by 3.3-fold. Furthermore, the cDNA clones of these CsPDAT genes were isolated for transient expression in tobacco leaves. All five genes showed PDAT enzymatic activity and substantially increased TAG accumulation in the leaves, with CsPDAT1-A showing a higher preference for ɑ-linolenic acid (18:3 ω-3). Overall, this study demonstrated that different members of CsPDAT family contribute to TAG synthesis in different tissues. More importantly, they are involved in different types of stress responses in camelina seedlings, providing new evidence of their roles in oil biosynthesis and regulation in camelina vegetative tissue. The identified CsPDATs may have practical applications in increasing oil accumulation and enhancing stress tolerance in other plants as well.
Myzus persicae and Bemisia tabaci are serious pests of tobacco and can occur simultaneously on the same plant. We found that tobacco plants infested by whiteflies had fewer aphids than those without whiteflies. To determine whether B. tabaci feeding could induce plant defense against aphids locally and systemically, we determined the effects of B. tabaci on several biological parameters of M. persicae on tobacco. Infestation of B. tabaci nymphs reduced survival rates of M. persicae by 30.0%. In three generations, M. persicae populations increased 1,091-fold on uninfested plants compared with 222-fold on the plants with whiteflies. On the upper leaves with systemic damage but uninfested B. tabaci, the survival rate of aphids was 9.3-fold lower than that on plants that were never been infested by whitefly. Survival rates of M. persicae on leaves with whiteflies present or with whiteflies removed were also lower than those on uninfested leaves. Fecundity of M. persicae was not different on leaves with whiteflies, with whiteflies removed or uninfested leaves; however, fecundity on leaves with systemic damage was lower than on uninfested leaves. Growth rates of M. persicae on the leaves with whiteflies, or with whiteflies removed, were higher than on uninfested leaves, whereas it was lower on systemically damaged leaves than on uninfested leaves. The development of M. persicae was approximately 1 d longer on systemic leaves with whiteflies than on uninfested leaves. These results indicate that feeding of B. tabaci induced a defense in tobacco plants against M. persicae, both locally and systemically, although other mechanisms may also be involved.
Palmitoleic acid (16:1Δ 9 ) is one kind of ω-7 fatty acids (ω-7 FAs) widely used in food, nutraceutical and industry. However, such high-valued ω-7 FA only has a trace level in mature seeds of cotton and other common oil crops. We found that palmitoleic acid (>10.58 Mol%) was specially enriched in developing cotton endosperm which is disappeared in its mature seed. The present study was conducted to investigate the mechanism underlying high accumulation of palmitoleic acid in developing endosperm but not in embryo of upland cotton ( Gossypium hirsutum L.) seed. Of 17 stearoyl-ACP Δ 9 desaturases (SAD) gene family members identified in upland cotton, six GhSADs may specifically work in the desaturation of palmitic acid (16:0-ACP) to produce palmitoleic acid (16:1Δ 9 -ACP), which were revealed by examining the key amino acids in the catalytic center and their cis -elements. Gene expression analysis showed that spatial patterns of these GhSADs were different in developing ovules, with GhA-SAD6 and GhD-SAD8 preferentially expressed in developing endosperms. Functional analysis by transient expression in Nicotiana benthamiana leaves and genetic complementary assay using yeast mutant BY4389 strain unable to synthesize unsaturated fatty acids demonstrated that GhA-SAD6 and GhD-SAD8 have strong substrate specificity for 16:0-ACP. In contrast, GhA-SAD5 and GhA-SAD7 exhibited high specific activity on 18:0-ACP. Taken together, these data evidence that GhA-SAD6 and GhD-SAD8 are responsible for making palmitoleic acid in developing cotton endosperms, and provide endogenous gene targets for genetic modification to enrich ω-7 FAs in cotton seed oil required for sustainable production of functionality-valued products.
Bradysia odoriphaga and Bradysia difformis are devastating pests of vegetable, ornamental crops and edible mushrooms causing significant losses. Temperature may be an important factor restricting their population abundance in the summer. To determine the effects of short-term heat shock on adults, their survival, longevity and fecundity data were collected, and antioxidant responses and heat shock protein expression levels were examined. Our results indicated that the survival rates of Bradysia adults decreased rapidly after heat shock ≥36 °C, and the longevity and reproductive capacities were significantly inhibited, indicating that short-term heat shock had lethal and sub-lethal effects. Moreover, the lipid peroxidation levels of B. difformis and B. odoriphaga increased dramatically at 36 °C and 38 °C, respectively. Four antioxidant enzymes activities of B. odoriphaga were greater than those of B. difformis at 38 °C. Additionally, hsp70 and hsp90 expression levels significantly increased after heat stress, and higher expression levels of B. difformis and B. odoriphaga were discovered at 36 and 38 °C respectively, indicating their different heat tolerance levels. Overall, short-term heat shock (≥36 °C) caused significantly adverse effects on Bradysia adults, indicating that it could be applied in pest control, and antioxidant system and hsp genes played important roles in their heat tolerance levels.
The plant-specific WRINKLED1 (WRI1) is a member of the AP2/EREBP class of transcription factors that positively regulate oil biosynthesis in plant tissues. Limited information is available for the role of WRI1 in oil biosynthesis in castor bean (Ricinus connunis L.), an important industrial oil crop. Here, we report the identification of two alternatively spliced transcripts of RcWRI1, designated as RcWRI1-A and RcWRI1-B. The open reading frames of RcWRI1-A (1341 bp) and RcWRI1-B (1332 bp) differ by a stretch of 9 bp, such that the predicted RcWRI1-B lacks the three amino acid residues “VYL” that are present in RcWRI1-A. The RcWRI1-A transcript is present in flowers, leaves, pericarps and developing seeds, while the RcWRI1-B mRNA is only detectable in developing seeds. When the two isoforms were individually introduced into an Arabidopsis wri1-1 loss-of-function mutant, total fatty acid content was almost restored to the wild-type level, and the percentage of the wrinkled seeds was largely reduced in the transgenic lines relative to the wri1-1 mutant line. Transient expression of each RcWRI1 splice isoform in N. benthamiana leaves upregulated the expression of the WRI1 target genes, and consequently increased the oil content by 4.3–4.9 fold when compared with the controls, and RcWRI1-B appeared to be more active than RcWRI1-A. Both RcWRI1-A and RcWRI1-B can be used as a key transcriptional regulator to enhance fatty acid and oil biosynthesis in leafy biomass.
The basic leucine-region zipper (bZIP) transcription factors (TFs) act as crucial regulators in various biological processes and stress responses in plants. Currently, bZIP family members and their functions remain elusive in the green unicellular algae Chlamydomonas reinhardtii, an important model organism for molecular investigation with genetic engineering aimed at increasing lipid yields for better biodiesel production. In this study, a total of 17 C. reinhardtii bZIP (CrebZIP) TFs containing typical bZIP structure were identified by a genome-wide analysis. Analysis of the CrebZIP protein physicochemical properties, phylogenetic tree, conserved domain, and secondary structure were conducted. CrebZIP gene structures and their chromosomal assignment were also analyzed. Physiological and photosynthetic characteristics of C. reinhardtii under salt stress were exhibited as lower cell growth and weaker photosynthesis, but increased lipid accumulation. Meanwhile, the expression profiles of six CrebZIP genes were induced to change significantly during salt stress, indicating that certain CrebZIPs may play important roles in mediating photosynthesis and lipid accumulation of microalgae in response to stresses. The present work provided a valuable foundation for functional dissection of CrebZIPs, benefiting the development of better strategies to engineer the regulatory network in microalgae for enhancing biofuel and biomass production.
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