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A New Species of Genus Microhyla (Amphibia: Anura: Microhylidae) from Zhejiang Province, China
Get PDFWe described a new species, Microhyla beilunensis sp. nov., from Zhejiang Province of China. Phylogenetic analyses based on the mitochondrial 12S, 16S and CO1 gene sequences suggested that the new taxon was distinctly separated from its congeners and closed to M. mixtura and M. okinavensis. Morphologically, the new species could be identified from its congeners except M. mixtura by several characters: (1) rudimentary webs on toe base; (2) absence of disks and dorsal median longitudinal grooves on finger tips; (3) presence of disks and dorsal median longitudinal grooves on toe tips. As well, the new species could be identified from topotype M. mixtura by the combination of characters: (1) apart from the stripes, bar-shaped and oval-shaped patterns, the rounded spots present on the dorsum of body and legs; (2) the outer metacarpal tubercles prominently larger than the inner one; (3) of males, the ratios of HW, IND, UEW and LAW to SVL of the new species were significantly larger than those of M. mixtura (P < 0.01), and the ratios of SL, IOD, LAHL, HLL, TL, TFL and FL to SVL of the new species were significantly less than those of M. mixtura (P < 0.05)
Population Size, Genetic Diversity and Molecular Evidence of a Recent Population Bottleneck in Hynobius chinensis, an Endangered Salamander Species
Get PDFSevere population declines can reduce species to small populations, offering permissive conditions for deleterious processes. For example, following such events, species can become prone to inbreeding and genetic drift which can lead to a loss of genetic diversity and evolutionary potentials. Hynobius chinensis is a poorly studied very rare and declining endangered amphibian species endemic to China in Changyang County. We investigated adult census population size by monitoring breeding populations from 2015 to 2018, developed microsatellite markers from the transcriptome and used them to investigate genetic diversity, and a population bottleneck in this species. We found H. chinensis in 4 different localities in a total area of 2.18 km2 and estimated the overall adult census population size at 386–404 individuals. The adult census size (mean ± SE) per breeding pond ranged from 44 ± 6 to 141 ± 8 individuals and appeared smaller than that reported in closely related species in undisturbed habitats. We developed and characterized 13 microsatellite markers in total. Analysis of data at 7 loci (N = 118) in Hardy-Weinberg equilibrium gathered from the largest population showed that genetic diversity level was low. The average number of alleles per locus was 2.14. The observed and expected heterozygosities averaged 0.38 and 0.40, respectively. The inbreeding coefficient was –0.06. All tests performed to investigate a population bottleneck, i.e. The Garza-Williamson test, Heterozygosity excess test, Mode shift test of allele frequency, and effective population size estimates detected a population bottleneck. The contemporary and the historical effective population sizes were estimated at 36 and 234 individuals, respectively. We argue that as bottleneck effects, the studied population may have become prone to genetic drift and inbreeding, losing microsatellite alleles and heterozygosity. Our results suggest that populations of H. chinensis may have been extirpated in the study area
A Review for Life-history Traits Variation in Frogs Especially for Anurans in China
Get PDFEnvironmental variation can promote differentiation in life-history traits in species of anurans. Increased environmental stress usually results in larger age at sexual maturity, older mean age, longer longevity, slower growth, larger body size, and a shift in reproductive allocation from offspring quantity to quality, and a stronger trade-off between offspring size and number. However, previous studies have suggested that there are inconsistent geographical variations in life-history traits among anuran species in China. Hence, we here review the intraspecific patterns and differences in life-history traits (i.e., egg size, clutch size, testes size, sperm length, age at sexual maturity, longevity, body size and sexual size dimorphism) among different populations within species along geographical gradients for anurans in China in recent years. We also provide future directions for studying difference in sperm performance between longer and shorter sperm within a species through transplant experiments and the relationships between metabolic rate and brain size and life-history
Perch Associated Expression of Phenotypic Plasticity in Limb Development and Sprint Speed in Agamid Lizard Calotes versicolor: A Laboratory Study
Get PDFNew born hatchlings of Calotes versicolor were reared in terrarium having narrow or wide perches for a period of 4-month and their snout vent length (SVL), tail, fore and hindlimb lengths were measured at monthly intervals. Limb postures (closer to the body or spread away from the body) were also recorded. The sprint speed was recorded in two and four-month old lizards on a 1 m long race track providing 45° or 60° slope. In both the groups, SVL and tail lengths were comparable but the limb lengths and their growth rates were significantly greater in lizards of wider perch group. The lizards reared with narrow perches positioned their limbs closer to the body; while those reared on wider perches spread their limbs away from their body. Further, the latter exhibited significantly higher sprint speed regardless of the slope of the race track over those of narrow perch group. Sprint speeds of lizards in both groups were correlated with the limb sizes. The study showed that the lizards reared on narrow or wide perches exhibited divergent adaptive responses (phenotypic plasticity) by developing longer or shorter limbs and corresponding changes in their sprint speeds. These findings support the idea that availability of perch structure during early development evokes adaptive plasticity in the limb development and associated locomotory performance in arboreal lizards like C. versicolor
Sexual Dimorphism, Female Reproductive Characteristics and Egg Incubation in an Oviparous Forest Skink (Sphenomorphus incognitus) from South China
Get PDFWe studied sexual dimorphism and female reproduction in an oviparous forest skink (Sphenomorphus incognitus) from South China. We incubated eggs under five thermal regimes (22, 25, 28, 25 ± 3 and 27 ± 5 °C) to examine the effects of constant versus fluctuating temperatures on incubation length and hatchling morphology. In our sample the largest male and female were 110 mm and 108 mm snout-vent length (SVL), respectively. Adult males and females did not differ in mean SVL; adult males were larger in head size (both length and width), longer in fore- and hind-limb lengths and shorter in abdomen length than females of the same SVL. Accordingly, we conclude that S. incognitus is a sexually monomorphic species in terms of SVL but shows sexual dimorphism in head size, abdomen length and appendage length. Females laid a single clutch of 3−10 eggs per breeding season from early May to mid-August, with larger females generally laying more (but not always larger) eggs per clutch than did smaller ones. Embryonic stages at laying ranged from Dufaure and Hubert’s (1961) stage 31 to 32, with a mean stage of 31.3. The positive relationship between clutch mass and female SVL was not significant. The offspring size-number trade-off does not exist in S. incognitus, as revealed by the fact that egg mass was independent of relative fecundity. Incubation length decreased as temperature increased, and stable temperatures resulted in delayed hatching. Hatchlings incubated under the five thermal regimes did not differ from each other in any examined trait, suggesting that S. incognitus is among oviparous reptilian species where incubation temperature has no role in modifying hatchling morphology as long as eggs are not exposed to extreme temperatures for prolonged periods of time
The Mitochondrial Genome of Pseudocalotes microlepis (Squamata: Agamidae) and its Phylogenetic Position in Agamids
Get PDFWe describe the complete mitochondrial genome of a small-scaled forest agamid (Pseudocalotes microlepis), which is 17 873 bp in size, containing 13 PCGs, 2 rRNAs, 22 tRNAs and non-coding regions. The mitogenome has a typical gene order among squamates. 13 PCGs include 2 start codons (ATG and ATA), 3 stop codons (TAG, TAA and AGG), and an incomplete stop codon (T-). Codon usage analyses showed that CUA-Leu1 and CGG-Arg are the most frequently and rarely used codon, respectively. All 22 tRNAs were predicted to form canonical cloverleaf secondary structures, except for two tRNAs (tRNACys and tRNASer (AGY)) lacking the dihydorouridine (DHU) arm. The large non-coding region (control region) is 2 687 bp long (28.3% T, 18.2% C, 42.3% A, and 11.2% G), with four different types of repeating sequences. The phylogenetic tree resulting from BEAST analyses based on concatenated 2 rRNAs and 13 PCGs in sequence revealed that the newly sequenced P. microlepis, where the genus Acanthosaura were aggregated. Together with Calotes versicolor, they constitute the subfamily Draconinae
Optimization of Vortex-Assisted Dispersive Liquid-Liquid Microextraction for the Simultaneous Quantitation of Eleven Non-Anthocyanin Polyphenols in Commercial Blueberry Using the Multi-Objective Response Surface Methodology and Desirability Function Approach
No full textIn the present study, 11 non-anthocyanin polyphenols, gallic acid, protocatechuate, vanillic acid, syringic acid, ferulic acid, quercetin, catechin, epicatechin, epigallocatechin gallate, gallocatechin gallate and epicatechin gallatewere firstly screened and identified from blueberries using an ultra performance liquid chromatography-time of flight mass spectrography (UPLC-TOF/MS) method. Then, a sample preparation method was developed based on vortex-assisted dispersive liquid-liquid microextraction. The microextraction conditions, including the amount of ethyl acetate, the amount of acetonitrile and the solution pH, were optimized through the multi-objective response surface methodology and desirability function approach. Finally, an ultra performance liquid chromatography-triple quadrupole mass spectrography (UPLC-QqQ/MS) method was developed to determine the 11 non-anthocyanin polyphenols in 25 commercial blueberry samples from Sichuan province and Chongqing city. The results show that this new method with high accuracy, good precision and simple operation characteristics, can be used to determine non-anthocyanin polyphenols in blueberries and is expected to be used in the analysis of other fruits and vegetables
Frond transformation system mediated by Agrobacterium tumefaciens for Lemna minor
No full textThe Lemnaceae, known as duckweed, the smallest flowering aquatic plant, shows promise as a plant bioreactor. For applying this potential plant bioreactor, establishing a stable and efficient genetic transformation system is necessary. The currently favored callus-based method for duckweed transformation is time consuming and genotype limited, as it requires callus culture and regeneration, which is inapplicable to many elite duckweed strains suitable for bioreactor exploitation. In this study, we attempted to establish a simple frond transformation system mediated by Agrobacterium tumefaciens for Lemna minor, one of the most widespread duckweed species in the world. To evaluate the feasibility of the new transformation system, the gene CYP710A11 was overexpressed to improve the yield of stigmasterol, which has multiple medicinal purposes. Three L. minor strains, ZH0055, D0158 and M0165, were transformed by both a conventional callus transformation system (CTS) and the simple frond transformation system (FTS). GUS staining, PCR, quantitative PCR and stigmasterol content detection showed that FTS can produce stable transgenic lines as well as CTS. Moreover, compared to CTS, FTS can avoid the genotype constraints of callus induction, thus saving at least half of the required processing time (CTS took 8-9 months while FTS took approximately 3 months in this study). Therefore, this transformation system is feasible in producing stable transgenic lines for a wide range of L. minor genotypes
Transcriptome analysis provides novel insights into high-soil-moisture-elevated susceptibility to Ralstonia solanacearum infection in ginger (Zingiber officinale Roscoe cv. Southwest)
No full textGinger (Zingiber officinale Roscoe), one of the most economically valuable plants in the Zingiberaceae family, is widely used as a spice and flavoring agent for beverages, bakery, confectionary, and pharmaceutics. Bacterial wilt disease, caused by Ralstonia solanacearum, is one of the most detrimental production constraints in ginger cultivation. Field cultivation experiments indicated that soil moisture affects the incidence of bacterial wilt disease. However, the relationship between soil moisture and bacterial wilt incidence as well as the mechanism that underlie this infection remain unclear. This study confirms that high soil moisture elevates the susceptibility to R. solanacearum infection; transcriptome sequencing was performed to elucidate the underlying mechanisms. Differential expression indicates that a small number of genes is involved in both the response to high soil moisture as well as post successful R. solanacearum infection; furthermore, a large number of genes is involved in the defense of the infection. In response to high soil moisture, higher ABA contents, and higher expression levels of ABF4 may be related to higher tiller density in ginger. More importantly, WAK16 and WAK3-2 may be determinative genes that weaken the resistance to R. solanacearum in ginger under high soil moisture. The down-regulated expression levels of PRX, CPY, and XET genes indicate that in response to successful R. solanacearum infection, the normal cell wall metabolism may be disturbed and the hypersensitive response may be inhibited. In summary, our study deepens our understanding of the molecular mechanisms of the soil moisture dependent wilt susceptibility of ginger
Water table drawdown reshapes soil physicochemical characteristics in Zoige peatlands
No full textWarming and human activities have caused peatland degradation and water table drawdown, which greatly influence the physicochemical characteristics of soil at different depths. How water table drawdown affects soil physicochemical characteristics is not well understood for the Tibetan Plateau, especially the Zoige peatlands. This study measured variations in C/N ratio, ash content, delta C-13, and humification index (HI) as a function of water table drawdown and depth by sampling 100-cm soil columns from three sites with different water tables in the Zoige peatlands. C/N ratio decreased while ash content and HI increased with water table drawdown, suggesting greater decomposition of soil organic matter at sites with a lower water table. The response of soil physicochemical characteristics to water table drawdown varied with depth: in the layer above the water table, C/N ratio did not vary, ash content decreased and HI increased; in the layer below the water table, C/N ratio and HI decreased, while ash content increased. The physicochemical characteristics above the water table were influenced mainly by new carbon input, while the characteristics below the water table were influenced mainly by decomposition. The depth profile of C/N ratio, ash content and HI aligned well with peat formation and decomposition and may be indicators of peat degradation. delta C-13 did not vary with water table drawdown, depth, or decomposition process. These insights into the variation of soil physicochemical characteristics with depths and water table may help elucidate the processes behind peatland degradation