Relationship between leaf vein traits of three dominant Quercus species and ecological factors in the Qinling Mountains, China.

We investigated the inter- and intra-species differences of leaf vein traits of three dominant Quercus species, Q. wutaishanica, Q. aliena var. acutiserrata, and Q. variabilis of Niubeiling (subtropical humid climate) and Taohuagou (warm temperate semi-humid climate), located in the eastern and western Qinling Mountains. The nine examined leaf vein traits included primary leaf vein width, secondary leaf vein width, mean fine vein width, primary vein density, fine vein density, vein areole diameter, areole density, 3D fine vein surface area, and fine vein volume. We further elucidated the influencing mechanisms and regulatory pathways of biotic and abiotic factors on leaf vein traits. The results showed that species identity had significant effects on eight out of nine leaf vein traits except 3D fine vein surface area, while habitat had significant effects on primary leaf vein width, secondary leaf vein width, vein areole diameter, fine vein density, and areole density. Altitude had significant effects on primary vein density, mean fine vein width, vein areole diameter, fine vein density and areole density. Habitat, tree species identity, and altitude had significantly interactive effects on primary leaf vein density, 3D fine vein surface area, and fine vein volume. There were significant differences in primary leaf vein width, mean fine vein width, areole density, 3D fine vein surface area, fine vein volume, primary vein density of Q. wutaishanica between the two studied habitats, but the differences were only found in secondary leaf vein width and areole density of Q. aliena var. acutiserrata and Q. variabilis. The examined leaf vein traits were influenced both by biotic and abiotic factors, with varying effect sizes. Among the biotic factors, petiole length, leaf length and width ratio had strong effect on leaf vein traits. Among the abiotic factors, climatic and soil factors had high effect size on vein traits, with the former being higher than the latter. Leaf vein traits were affected directly by biotic factors, but indirectly by abiotic factors (soil and climatic factors) via regulating biotic factors (leaf stoichiometry and leaf phenotypic traits).

Comparison of species composition and community characteristics of Quercus forests on south and north slopes of Taibai Mountain, China.

We examined species composition, community characteristics, diversity, and community similarity of five Quercus communities composed of three Quercus species (Q. variabilis, Q. aliena var. acutiserrata, Q. wutaishanica) on the altitudinal gradient on the south and north slopes of Taibai Mountain. The results showed that there was an altitudinal transition pattern from Q. variabilis pure forest to Q. variabilis-Q. aliena var. acutiserrata mixed forest, Q. aliena var. acutiserrata pure forest, Q. aliena var. acutiserrata-Q. wutaishanica mixed forest and Q. wutaishanica pure forest on the south and north slopes of Taibai Mountain. The main companion species of Quercus community on the north slope were Pinus armandii, Castanea seguinii, and Sorbus alnifolia, and were Pinus tabuliformis, C. seguinii, Carpinus cordata, and Q. spinosa on the south slope. Species richness, woody plant density, and Quercus species dominance on the north slope of Taibai Mountain were higher than those on the south slope. α diversity of tree layer in Quercus community on the south and north slopes of Taibai Mountain increased first, then decreased and then increased with altitude. α diversity of tree layer was higher in mixed forests than pure forests. α diversity of shrub layer was higher than that of tree layer and herb layer in Quercus community on south and north slopes. ß diversity fluctuated greatly along the altitudinal gradient on the south and north slopes, indicating that species composition changed greatly with altitude. Results of redundancy analysis showed that mean warmest month temperature, altitude and tree height accounted for 79.0% of the community diversity on the north slope, and that soil water content, tree height, canopy density and mean annual temperature accounted for 79.6% of the community diversity on the south slope. Overall, Quercus dominance was higher on the north slope of Taibai Mountain, and the substitution distribution pattern of Quercus species was clearer than that on the south slope. Environmental factors related to temperature and precipitation jointly affected α diversity of Quercus communities.

[Expression, purification and characterization of a novel bis (hydroxyethyl) terephthalate hydrolase from Hydrogenobacter thermophilus].

PET (polyethylene terephthalate) is one of the most important petrochemicals that is widely used in mineral water bottles, food and beverage packaging and textile industry. Because of its stability under environmental conditions, the massive amount of PET wastes caused serious environmental pollution. The use of enzymes to depolymerize PET wastes and upcycling is one of the important directions for plastics pollution control, among which the key is the depolymerization efficiency of PET by PET hydrolase. BHET (bis(hydroxyethyl) terephthalate) is the main intermediate of PET hydrolysis, its accumulation can hinder the degradation efficiency of PET hydrolase significantly, and the synergistic use of PET hydrolase and BHET hydrolase can improve the PET hydrolysis efficiency. In this study, a dienolactone hydrolase from Hydrogenobacter thermophilus which can degrade BHET (HtBHETase) was identified. After heterologous expression in Escherichia coli and purification, the enzymatic properties of HtBHETase were studied. HtBHETase shows higher catalytic activity towards esters with short carbon chains such as p-nitrophenol acetate. The optimal pH and temperature of the reaction with BHET were 5.0 and 55 ℃, respectively. HtBHETase exhibited excellent thermostability, and retained over 80% residual activity after treatment at 80 ℃ for 1 hour. These results indicate that HtBHETase has potential in biological PET depolymerization, which may facilitate the enzymatic degradation of PET.

[Engineering the plastic degradation enzyme Ple629 from marine consortium to improve its thermal stability].

Petrochemical-derived polyester plastics such as polyethylene terephthalate (PET) and polybutylene adipate terephthalate (PBAT) have been widely used. However, the difficulty to be degraded in nature (PET) or the long biodegradation cycle (PBAT) resulted in serious environmental pollution. In this connection, treating these plastic wastes properly becomes one of the challenges of environment protection. From the perspective of circular economy, biologically depolymerizing the waste of polyester plastics and reusing the depolymerized products is one of the most promising directions. Recent years have seen many reports on polyester plastics degrading organisms and enzymes. Highly efficient degrading enzymes, especially those with better thermal stability, will be conducive to their application. The mesophilic plastic-degrading enzyme Ple629 from the marine microbial metagenome is capable of degrading PET and PBAT at room temperature, but it cannot tolerate high temperature, which hampers its potential application. On the basis of the three-dimensional structure of Ple629 obtained from our previous study, we identified some sites which might be important for its thermal stability by structural comparison and mutation energy analysis. We carried out transformation design, and performed expression, purification and thermal stability determination of the mutants. The melting temperature (Tm) values of mutants V80C and D226C/S281C were increased by 5.2 ℃ and 6.9 ℃, respectively, and the activity of mutant D226C/S281C was also increased by 1.5 times compared with that of the wild-type enzyme. These results provide useful information for future engineering and application of Ple629 in polyester plastic degradation.

Molecular and Biochemical Differences of the Tandem and Cold-Adapted PET Hydrolases Ple628 and Ple629, Isolated From a Marine Microbial Consortium.

Polybutylene adipate terephthalate (PBAT) is a biodegradable alternative to polyethylene and can be broadly used in various applications. These polymers can be degraded by hydrolases of terrestrial and aquatic origin. In a previous study, we identified tandem PETase-like hydrolases (Ples) from the marine microbial consortium I1 that were highly expressed when a PBAT blend was supplied as the only carbon source. In this study, the tandem Ples, Ple628 and Ple629, were recombinantly expressed and characterized. Both enzymes are mesophilic and active on a wide range of oligomers. The activities of the Ples differed greatly when model substrates, PBAT-modified polymers or PET nanoparticles were supplied. Ple629 was always more active than Ple628. Crystal structures of Ple628 and Ple629 revealed a structural similarity to other PETases and can be classified as member of the PETases IIa subclass, α/ß hydrolase superfamily. Our results show that the predicted functions of Ple628 and Ple629 agree with the bioinformatic predictions, and these enzymes play a significant role in the plastic degradation by the consortium.

Structural insight and engineering of a plastic degrading hydrolase Ple629.

Polyethylene terephthalate (PET) is one of the most abundantly produced synthetic polyesters. The vast number of waste plastics including PET has challenged the waste management sector while also posing a serious threat to the environment due to improper littering. Recently, enzymatic PET degradation has been shown to be a viable option for a circular plastic economy, which can mitigate the plastic pollution. While protein engineering studies on specific PET degradation enzymes such as leaf-branch compost cutinase (LCC), Thermobifida sp. cutinases and Ideonella sakaiensis PETase (IsPETase) have been extensively published, other homologous PET degrading enzymes have received less attention. Ple629 is a polyester hydrolase identified from marine microbial consortium having activity on PET and the bioplastic polybutylene adipate terephthalate (PBAT). In order to explore its catalytic mechanism and improve its potential for PET hydrolysis, we solved its crystal structure in complex with a PET monomer analogue, and validated its structural and mechanistic similarity to known PET hydrolases. By structural comparisons, we identified some hot spot positions described in previous research on protein engineering of PET hydrolases. We substitute these amino acid residues in Ple629, and obtained variants with improved activity and thermo-stability. The most promising variant D226A/S279A exhibited a more than 5.5-fold improved activity on PET nanoparticles than the wild-type enzyme, suggesting its potential applicability in the biotechnological plastic recycling.

Parasitism of Two Spodoptera spp. by Microplitis prodeniae (Hymenoptera: Braconidae).

Early instar larvae of the tobacco cutworm Spodoptera litura (Lepidoptera: Noctuidae) and the beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) are recognized hosts of the parasitic wasp Microplitis prodeniae Rao and Kurian (Hymenoptera: Braconidae), although M. prodeniae has previously been regarded as monophagous. In this study, we found the immature period and longevity of M. prodeniae developing in S. exigua was similar to that in S. litura. It was shown that the development time of M. prodeniae in S. exigua was 15.1 ± 0.3 d, not significantly different from 15.0 ± 0.2 d in S. litura. The parasitism rate of M. prodeniae attacking S. exigua was significantly lower than on S. litura (65.48 ± 2.29 and 43.83 ± 2.20%, respectively), whilst the female ratio of the wasp's offspring was not significantly different when developing on the two species. M. prodeniae females prefer to oviposit on the second- and third-instar host larvae of S. exigua, rather than other instars. The effects of development of M. prodeniae on two important lepidopterous pests are discussed.

A Place to Grow? Host Choice and Larval Performance of Microplitis similis (Hymenoptera: Braconidae) in the Host Spodoptera litura (Lepidoptera: Noctuidae).

Host selection is a key stage in the lifecycle of parasitoids, and is critical to both their function in control and to the maintenance of their population. The solitary endoparasitoid Microplitis similis (Hymenoptera: Braconidae) is a potential biological control agent of Spodoptera litura (F.) larvae (Lepidoptera: Noctuidae). In this study, we examined the preference M. similis exhibits for different instars of the host, host instar effects on parasitoid development, and the weight gain and food consumption of different instars of parasitized larvae. In no-choice tests, parasitization rates were highest in second- and early third-instar larvae, and no fourth- or fifth-instar hosts were parasitized. When provided with a choice of first- to late third-instar host larvae, M. similis preferred to parasitize early third-instar host larvae (41%) with a selection coefficient of 0.37. All morphometric features of wasp offspring increased with increasing age of the host at parasitization. A lower proportion of females emerged from first-instar larvae than any other instar. Parasitized S. litura larvae showed a pronounced reduction in food consumption and weight gain. Microplitis similis may have the potential to significantly suppress population growth and the damage caused by S. litura.

Imperfection works: Survival, transmission and persistence in the system of Heliothis virescens ascovirus 3h (HvAV-3h), Microplitis similis and Spodoptera exigua.

Ascoviruses are insect-specific large DNA viruses that mainly infect noctuid larvae, and are transmitted by parasitoids in the fields. Heliothis virescens ascovirus 3h (HvAV-3h) has been recently isolated from Spodoptera exigua, without parasitoid vector identified previously. Here we report that Microplitis similis, a solitary endoparasitoid wasp, could transmit HvAV-3h between S. exigua larvae in the laboratory. When the female parasitoid wasp acquired the virus and served as a vector, the period of virion viability on the ovipositor was 4.1 ± 1.4 days. Infected host larvae were still acceptable for egg laying by parasitoids, and the parasitoids thereafter transmitted virus to healthy hosts. Virus acquisition occurred only from donor hosts between 3 and 9 days post infection. The peak of virus acquisition (80.9 ± 6.3%) was found when M. similis wasps oviposited in larvae that had been inoculated with the virus 7 days previously. When virus infection of the host took place during the life cycle of the parasitoid wasp, it caused 1- to 4-day-old immature parasitoids death in the host, whilst a small proportion of 5- to 6-day-old and the majority of 7-day-old parasitoids larvae survived from the virus-infected hosts. Viral contamination did not reduce the life span or fecundity of female M. similis.

A comparison of growth and development of three major agricultural insect pests infected with Heliothis virescens ascovirus 3h (HvAV-3h).

Ascoviruses are double-stranded DNA viruses that are pathogenic to lepidopteran hosts, particularly noctuid larvae. Infection of a larva is characterized by retarded growth, reduced feeding and yellowish body color. In this paper, we reported the growth and development of three major agricultural noctuid insect pests, Helicoverpa armigera (Hübner), Spodoptera exigua (Hübner) and Spodoptera litura (Fabricius), infected with Heliothis virescens ascovirus 3h (HvAV-3h). Using 10-fold serial dilutions (0 to 7) of HvAV-3h-containing hemolymph to infect S. litura larvae, we found no significant difference in larval mortalities from 0 to 10(3)-fold dilutions; however, significant differences were observed at 10(4)-fold dilution and above. Using a 10-fold dilution of HvAV-3h-containing hemolymph to infect H. armigera, S. exigua and S. litura larvae, we found that the growth and development were significantly affected. All infected larvae could not pupate; the survival times of treated H. armigera, S. litura and S. exigua larvae were significantly longer than untreated control larvae. Body weight showed significant difference between treated and untreated control group from day 1 after inoculation in H. armigera and S. exigua, but day 2 in S. litura. Additionally, food intake also showed significant difference between treated and untreated control group from day 2 after inoculation in H. armigera and S. litura, but day 3 in S. exigua.