Genome-wide association studies reveal novel QTLs for agronomic traits in soybean.

Introduction: Soybean, as a globally significant crop, has garnered substantial attention due to its agricultural importance. The utilization of molecular approaches to enhance grain yield in soybean has gained popularity. Methods: In this study, we conducted a genome-wide association study (GWAS) using 156 Chinese soybean accessions over a two-year period. We employed the general linear model (GLM) and the mixed linear model (MLM) to analyze three agronomic traits: pod number, grain number, and grain weight. Results: Our findings revealed significant associations between qgPNpP-98, qgGNpP-89 and qgHGW-85 QTLs and pod number, grain number, and grain weight, respectively. These QTLs were identified on chromosome 16, a region spanning 413171bp exhibited associations with all three traits. Discussion: These QTL markers identified in this study hold potential for improving yield and agronomic traits through marker-assisted selection and genomic selection in breeding programs.

Effects of depth of straw returning on maize yield potential and greenhouse gas emissions.

Appropriate straw incorporation has ample agronomic and environmental benefits, but most studies are limited to straw mulching or application on the soil surface. To determine the effect of depth of straw incorporation on the crop yield, soil organic carbon (SOC), total nitrogen (TN) and greenhouse gas emission, a total of 4 treatments were set up in this study, which comprised no straw returning (CK), straw returning at 15 cm (S15), straw returning at 25 cm (S25) and straw returning at 40 cm (S40). The results showed that straw incorporation significantly increased SOC, TN and C:N ratio. Compared with CK treatments, substantial increases in the grain yield (by 4.17~5.49% for S15 and 6.64~10.06% for S25) were observed under S15 and S25 treatments. S15 and S25 could significantly improve the carbon and nitrogen status of the 0-40 cm soil layer, thereby increased maize yield. The results showed that the maize yield was closely related to the soil carbon and nitrogen index of the 0-40 cm soil layer. In order to further evaluate the environmental benefits of straw returning, this study measured the global warming potential (GWP) and greenhouse gas emission intensity (GHGI). Compared with CK treatments, the GWP of S15, S25 and S40 treatments was increased by 9.35~20.37%, 4.27~7.67% and 0.72~6.14%, respectively, among which the S15 treatment contributed the most to the GWP of farmland. GHGI is an evaluation index of low-carbon agriculture at this stage, which takes into account both crop yield and global warming potential. In this study, GHGI showed a different trend from GWP. Compared with CK treatments, the S25 treatments had no significant difference in 2020, and decreased significantly in 2021 and 2022. This is due to the combined effect of maize yield and cumulative greenhouse gas emissions, indicating that the appropriate straw returning method can not only reduce the intensity of greenhouse gas emissions but also improve soil productivity and enhance the carbon sequestration effect of farmland soil, which is an ideal soil improvement and fertilization measure.

Effects of salt stress on soil enzyme activities and rhizosphere microbial structure in salt-tolerant and -sensitive soybean.

Salt is recognized as one of the most major factors that limits soybean yield in acidic soils. Soil enzyme activity and bacterial community have a critical function in improving the tolerance to soybean. Our aim was to assess the activities of soil enzyme, the structure of bacteria and their potential functions for salt resistance between Salt-tolerant (Salt-T) and -sensitive (Salt-S) soybean genotypes when subject to salt stress. Plant biomass, soil physicochemical properties, soil catalase, urease, sucrase, amylase, and acid phosphatase activities, and rhizosphere microbial characteristics were investigated in Salt-T and Salt-S soybean genotypes under salt stress with a pot experiment. Salt stress significantly decreased the soil enzyme activities and changed the rhizosphere microbial structure in a genotype-dependent manner. In addition, 46 ASVs which were enriched in the Salt-T geotype under the salt stress, such as ASV19 (Alicyclobacillus), ASV132 (Tumebacillus), ASV1760 (Mycobacterium) and ASV1357 (Bacillus), which may enhance the tolerance to soybean under salt stress. Moreover, the network structure of Salt-T soybean was simplified by salt stress, which may result in soil bacterial communities being susceptible to external factors. Salt stress altered the strength of soil enzyme activities and the assembly of microbial structure in Salt-T and Salt-S soybean genotypes. Na+, NO3--N, NH4+-N and Olsen-P were the most important driving factors in the structure of bacterial community in both genotypes. Salt-T genotypes enriched several microorganisms that contributed to enhance salt tolerance in soybeans, such as Alicyclobacillus, Tumebacillus, and Bacillus. Nevertheless, the simplified network structure of salt-T genotype due to salt stress may render its bacterial community structure unstable and susceptible.

Salt altered rhizosphere fungal community and induced soybean recruit specific species to ameliorate salt stress.

Different crop genotypes showed different adaptability to salt stress, which is partly attributable to the microorganisms in the rhizosphere. Yet, knowledge about how fungal communities of different genotypes in soybean respond to salt stress is limited. Here, qPCR and ITS sequencing were used to assess the response of rhizobial fungal communities of resistant and susceptible soybean to salt stress. Moreover, we isolated two fungal species recruited by resistant soybeans for validation. The assembly of fungal community structure might be strongly linked to alterations in fungal abundance and soil physicochemical properties. Salt stress derived structural differences in fungal communities of resistant and susceptible genotypes. The salt-resistant genotype appeared to recruit some fungal taxa to the rhizosphere to help mitigating salt stress. An increase of fungal taxa with predicted saprotrophic lifestyles might help promoting plant growth by increasing nutrient availability to the plants. Compared with the susceptible genotypes, the resistant genotypes had more stronger network structure of fungi. Lastly, we verified that recruited fungi, such as Penicillium and Aspergillus, can soybean adapt to salt stress. This study provided a promising approach for rhizospheric fungal community to enhance salt tolerance of soybean from the perspective of microbiology and ecology.

Evolutionary Shift of Insect Diapause Strategy in a Warming Climate: An Intra-Population Evidence from Asian Corn Borer.

Herbivorous insects having variable numbers of generations annually depending on climate and day length conditions are increasingly breeding additional generations driven by elevated temperature under the scenario of global warming, which will increase insect abundance and result in more frequent damage events. Theoretically, this relies on two premises, i.e., either an evolutionary shift to facultative diapause for an insect behaving an obligatory diapause or developmental plasticity to alter voltinism productively for an insect with facultative diapause before shortening photoperiods inducing diapause. Inter-population evidence supporting the premise (theory) comes primarily from a model system with voltinism linked to thermal gradients across latitude. We examined the intra-population evidence in the field (47°24' N, 123°68' E) with Ostrinia furnacalis, one of the most destructive pests, on corn in Asia and Pacific islands. The species was univoltine in high latitudinal areas (≤46° N). Divergence of the diapause feature (obligatory and facultative) was observed within the field populations from 2016 to 2021. Warmer climates would provoke more facultative diapause individuals to initiate a second generation, which will significantly drive the population to evolve toward facultative diapause (multi-voltinism). Both divergent diapause and temperature must be considered for accurate prediction of phenology and population dynamics in ACB.

Comparison of Adverse Events between Isolated Left Atrial Appendage Closure and Combined Catheter Ablation.

(1) Background: This study aimed to investigate the effect of an additional catheter ablation (CA) procedure on the risk of post-procedure adverse events during CA combined with left atrial appendage closure (LAAC). (2) Methods: From July 2017 to February 2022, data from 361 patients with atrial fibrillation who underwent LAAC at our center were analyzed retrospectively. The adverse events were compared between CA + LAAC and LAAC-only groups. (3) Results: The incidence of device-related thrombus (DRT) and embolic events was significantly lower in the CA + LAAC group than in the LAAC-only group (p = 0.01 and 0.04, respectively). A logistic regression analysis revealed that the combined procedure served as a protective factor for DRT (OR = 0.09; 95% confidence interval: 0.01-0.89; p = 0.04). Based on a Cox regression analysis, the risk of embolism marginally increased in patients aged ≥65 years (HR = 7.49, 95% CI: 0.85-66.22 p = 0.07), whereas the combined procedure was found to be a protective factor (HR = 0.25, 95% CI: 0.07-0.87 p = 0.03). Further subgroup and interaction analyses revealed similar results. (4) Conclusions: The combined procedure may be associated with a lower rate of post-procedure DRT and embolization without a higher occurrence of other adverse events after LAAC. A risk-score-based prediction model was conducted, showing a good prediction performance.

Seven Years of Monitoring Susceptibility to Cry1Ab and Cry1F in Asian Corn Borer.

Resistance monitoring in the Asian corn borer, Ostrinia furnacalis, is necessary to accommodate the commercial introduction and stewardship of Bt maize in China. The susceptibility of 56 O. furnacalis field populations, collected between 2015 and 2021 from the corn belt regions of China, to Cry1Ab and Cry1F toxins was determined. Neonate larvae (within 12 h after hatching) were placed on the surface of semi-artificial agar-free diet incorporating a series of concentrations of purified toxins, and mortality was evaluated after 7d. The median lethal concentration (LC50) values of Cry1Ab and Cry1F were 0.05 to 0.37 µg/g (protein/diet) and 0.10 to 1.22 µg/g, respectively. Although interpopulation variation in susceptibility to the toxins was observed, the magnitude of the differences was 5.8-fold and 8.3-fold for Cry1Ab and Cry1F, respectively. These results suggested that the observed susceptibility differences reflect natural geographical variation in response and not variation caused by prior exposure to selection pressures. Therefore, the O. furnacalis populations were apparently still susceptible to Cry1Ab and Cry1F across their range within China. The monitoring data established here will serve as a comparative reference for early warning signs of field-evolved resistance after the cultivation of Bt maize in China.

LncRNA MDRL Mitigates Atherosclerosis through miR-361/SQSTM1/NLRP3 Signaling.

Objective: Long non-coding RNAs (lncRNAs) play many important roles in gene regulation and disease pathogenesis. Here, we sought to determine that mitochondrial dynamic related lncRNA (MDRL) modulates NLRP3 inflammasome activation and apoptosis of vascular smooth muscle cells (VSMCs) and protects arteries against atherosclerosis. Methods: In vivo experiments, we applied LDLR knockout (LDLR-/-) mice fed the high-fat diet to investigate the effects of MDRL on atherosclerosis. In vitro experiments, we applied mouse aortic smooth muscle cells to determine the mechanism of MDRL in abrogating NLRP3 inflammasome and inhibiting cell apoptosis through miR-361/sequentosome 1 (SQSTM1) by TUNEL staining, quantitative RT-PCR, western blot, microribonucleoprotein immunoprecipitation, and luciferase reporter assay. Results: Downregulated MDRL and increased NLRP3 were observed in mouse atherosclerotic plaques, accompanied with the increase of miR-361. The results showed that MDRL overexpression significantly attenuated the burden of atherosclerotic plaque and facilitated plaque stability through inhibiting NLRP3 inflammasome activation and cell apoptosis, and vice versa. Mechanically, MDRL suppressed NLRP3 inflammasome activation and VSMC apoptosis via suppressing miR-361. Furthermore, miR-361 directly bound to the 3'UTR of SQSTM1 and inhibited its translation, subsequently activating NLRP3 inflammasome. Systematic delivery of miR-361 partly counteracted the beneficial effects of MDRL overexpression on atherosclerotic development in LDLR-/- mice. Conclusions: In summary, MDRL alleviates NLRP3 inflammasome activation and apoptosis in VSMCs through miR-361/SQSTM1/NLRP3 pathway during atherogenesis. These data indicate that MDRL and inhibition of miR-361 represent potential therapeutic targets in atherosclerosis-related diseases.

The effect of different preoperative left atrial appendage emptying speeds on left atrial function in patients with persistent atrial fibrillation after left atrial appendage closure combined with catheter ablation.

OBJECTIVE: The present study aimed to investigate whether different preoperative left atrial appendage emptying speeds (LAAEVs) have an effect on left atrial function in patients with sinus arrhythmia after left atrial appendage closure (LAAC) combined with catheter ablation (CA). METHODS: A total of 163 patients with persistent non-valvular atrial fibrillation who received combined LAAC+CA surgery were included in the present study. Regular follow-up was conducted for 12 months, and patients with complete data and successful sinus rhythm recovery were selected as the study subjects (n = 82). The patients were divided into two groups: the LAAEV < 25 cm/s group and the LAAEV ≥ 25 cm/s group (n = 41 each). The propensity score was used for matching according to gender, age, CHA2DS2-VASc score, and HAS-BLED score. The changes in the two groups in LA structure, storage function, conduit function, and pump function were compared. RESULTS: Before surgery, the maximum LA volume (LAVmax) and minimum LA volume (LAVmin) were greater in the LAAEV < 25 cm/s group than in the LAAEV ≥ 25 cm/s group. The LA storage function (eg. Ƹ and SRs), conduit function (eg. SRe), and pump function (eg. SRa) were all worse in the LAAEV < 25 cm/s group than in the LAAEV ≥ 25 cm/s group. After the combined LAAC+CA surgery, the LA storage, conduit, and pump functions improved in both groups. At 12 months after surgery, there were no statistically significant differences between the two groups. CONCLUSION: Before combined LAAC+CA surgery, the LA structure and function of the LAAEV < 25 cm/s group were worse than those of the LAAEV ≥ 25 cm/s group. However, after LAAC+CA surgery, the LA structure and function of the patients were improved, and there were no significant differences between the two groups. Inferred improvement in LA structure and function in the LAAEV < 25 cm/s group was superior to that in the LAAEV ≥ 25 cm/s group.

First report of Corn Ear Rot Caused by Fusarium asiaticum in China.

Corn (Zea mays L.) ear rot, caused by various pathogens, is one of the most significant diseases of corn worldwide. In September 2020, a survey was undertaken to identify pathogenic fungi associated with corn ear rot in Suihua city (46.63°N 126.98°E), Heilongjiang Province, China. The average disease incidence was 14.2% and 15.6% in each of two fields sampled (~5 ha) using a five-point method (100 plants/each point). Twenty tissue samples from 20 diseased ears, showing white or pink mold on the surface of corn ears, were surface disinfected in 0.5% NaOCl for 5 min, rinsed 3 times in autoclaved distilled water. After drying, four treated corn kernels (one kernel/each ear) were placed onto potato dextrose agar (PDA) amended with 50 µg/mL streptomycin. The plates were sealed with parafilm sealing film and cultured in the dark at 26℃ with 80% RH for 3 days in an incubator. A total of 12 morphologically similar fungal isolates were obtained and subcultured by transferring hyphal tips for 3-5 days. Single-conidium isolates were generated with methods reported previously (Leslie and Summerell 2006). Colonies on PDA, reaching 20.3-20.9 mm·d-1 at 26℃, consisted of white to pale yellow, locculent, lush and dense aerial mycelium with red to apricot color. Macroconidia of six isolates randomly selected on carnation leaf agar (CLA) were falciform with a foot cell, three- to six-septate, measuring 12.6-67.2 × 2.6-5.4 µm (n = 100) in the dark at 26℃ with 80% RH for 5 days. No microconidia were observed. Based on these characteristics, the isolates were preliminarily identified as Fusarium asiaticum (Chang et al. 2020; Leslie and Summerell 2006). Genomic DNA of three representative isolates YSF2, YSF4 and YSF7 were extracted and the translation elongation factor 1-α (TEF-1ɑ) gene was amplified and sequenced using the primers EF1-728F/EF1-986R (Carbone and Kohn 1999). The DNA sequences of YSF2, YSF4 and YSF7 were deposited in GenBank (OL631287.1, OP272129 and OP272130). Analysis of TEF-1ɑ sequences of YSF2, YSF4 and YSF7 showed that they were 100% identical to F. asiaticum isolates NRRL 26156 (AF212452.1) in NCBI and NRRL 13818 (AF212451.1) in Fusarium MLST. A pathogenicity test was performed on corn cv. Xinxin 1. Four days after silk emergence, 3 mL conidial suspension (106 macroconidia/ml) of each 12 isolates was individually injected into the center of the ear through the husk sideways, penetrating the kernels to a depth of about 5 mm (6 ears/each isolate) in the field (Guo et al. 2020). Six corn ears treated with sterile distilled water were used as the control. After inoculation, normal field management was carried out. All inoculated ears showed symptoms similar to those observed in the field 50 days after inoculation, while no symptoms were observed on the blank control ears. Five fungal isolates with the same colony morphology as the naturally occurring ear rot in the field were re-isolated from the inoculated corn kernels and confirmed to be F. asiaticum according to morphological characteristics and sequence analysis of five fungal isolates. F. asiaticum has previously been reported to cause corn ear rots in Japan (Kawakami et al. 2015). To our knowledge, this is the first report of F. asiaticum causing corn ear rot in Northeast China. Corn ear rot poses a threat to significantly reduce the quality of corn in a major production zone of maize in China. Therefore, its distribution needs to be investigated and effective disease management strategies developed.

The response of grain yield and ear differentiation related traits to nitrogen levels in maize varieties with different nitrogen efficiency.

Maize (Zea mays L.) is one of the most widely distributed and important crops in China. Maize ear differentiation plays an important role grain yield formation. However, it is unclear if ear and root morphophysiology status affects yield formation by altering ear differentiation and development under different nitrogen (N) conditions. The aim of this study is to understand how the ear differentiation and development are affected by ear and root morphophysiology traits, as affected by the N rate. The experiment consisted of two N rates: high nitrogen (180 kg ha-1), and low nitrogen (60 kg ha-1). Two N-efficient varieties (NEVs) and two N-inefficient varieties (NIVs) were grown in the field. The results showed higher nitrogen accumulation and grain yield in NEVs than in NIVs, which was mainly attributed to the increased N uptake by the larger root system under both N conditions. Under high N conditions, among ear differentiation-related traits, only FR was significantly positively correlated with grain yield, and NEVs ensure FR through higher N concentration and ZR content in ear at the fertilization stage. Under low N conditions, NEVs obtained higher FP, SR and FR through higher N concentration and IAA in ear at the early stage of ear differentiation, maintained lower AR and BTL by higher RA, R-ZR and E-ZR at the late stage of ear growth. These results suggest that NEVs have a more complex mechanism for obtaining higher grain yield under low N conditions than N sufficiency, and that phytohormones play an important role in this process.

The Long-Term Results of Three Catheter Ablation Methods in Patients With Paroxysmal Atrial Fibrillation: A 4-Year Follow-Up Study.

Aims: Catheter ablation of paroxysmal atrial fibrillation (PAF) has been shown to be effective and safe. However, recurrence of PAF varies between 10 and 30% for radiofrequency ablation. There have been no reports comparing long-term recurrence rates following radiofrequency ablation, cryoablation, and three-dimensional guided cryoablation plus radiofrequency ablation. The aim of this study was to observe the long-term recurrence rate of PAF when treated by these three catheter ablation methods, and to explore clinical factors that can potentially predict PAF recurrence following catheter ablation. Methods: There were 238 patients involved in this study, including 106 radiofrequency (RF) ablation cases (RF group), 66 cryoablation cases (Freeze group), and 66 cases treated by three-dimensional guided cryoablation combined with radiofrequency ablation (Freeze-plus-RF group). All patients underwent standardized follow-up. The recurrence rate of atrial fibrillation (AF) in the three groups was calculated. Predictive factors for the recurrence of AF were also investigated. Results: At 48 months (the median follow-up period), the sinus rhythm maintenance rate was 77.4% in the RF group, 72.7% in the Freeze group, and 81.8% in the Freeze-plus-RF group. The maintenance rate of sinus rhythm was highest in the Freeze-plus-RF group, but differences among the three groups were not statistically significant. Further analysis found that the preoperative left atrial appendage emptying velocity (LAAEV) (recurrence vs. no recurrence, 56.58 ± 18.37 vs. 65.59 ± 18.83, respectively, p = 0.003), left atrial (LA) anteroposterior dimension (recurrence vs. no recurrence, 36.56 ± 4.65 vs. 35.00 ± 4.37, respectively; p = 0.028), and LA vertical dimension (recurrence vs. no recurrence, 56.31 ± 6.96 vs. 53.72 ± 6.52, respectively; p = 0.035) were related to postoperative recurrence. Multiple Cox regression analysis showed that only LAAEV was predictive of postoperative recurrence of PAF (hazard ratio: 0.979; 95% confidence interval: 0.961-0.997). Conclusion: Our study found that there was no statistically significant difference in long-term recurrence rates among the RF, Freeze, and Freeze-plus-RF groups. Preoperative LAAEV is an independent predictor of postoperative recurrence of PAF.

Rhizosphere Soil Bacterial Communities of Continuous Cropping-Tolerant and Sensitive Soybean Genotypes Respond Differently to Long-Term Continuous Cropping in Mollisols.

The continuous planting of soybeans leads to soil acidification, aggravation of soil-borne diseases, reduction in soil enzyme activity, and accumulation of toxins in the soil. Microorganisms in the rhizosphere play a very important role in maintaining the sustainability of the soil ecosystem and plant health. In this study, two soybean genotypes, one bred for continuous cropping and the other not, were grown in a Mollisol in northeast China under continuous cropping for 7 and 36years in comparison with soybean-maize rotation, and microbial communities in the rhizosphere composition were assessed using high-throughput sequencing technology. The results showed that short- or long-term continuous cropping had no significant effect on the rhizosphere soil bacterial alpha diversity. Short-term continuous planting increased the number of soybean cyst nematode (Heterodera glycines), while long-term continuous planting reduced these numbers. There were less soybean cyst nematodes in the rhizosphere of the tolerant genotypes than sensitive genotypes. In addition, continuous cropping significantly increased the potential beneficial bacterial populations, such as Pseudoxanthomonas, Nitrospira, and Streptomyces compared to rotation and short-term continuous cropping, suggesting that long-term continuous cropping of soybean shifts the microbial community toward a healthy crop rotation system. Soybean genotypes that are tolerant to soybean might recruit some microorganisms that enhance the resistance of soybeans to long-term continuous cropping. Moreover, the network of the two genotypes responded differently to continuous cropping. The tolerant genotype responded positively to continuous cropping, while for the sensitive genotype, topology analyses on the instability of microbial community in the rhizosphere suggested that short periods of continuous planting can have a detrimental effect on microbial community stability, although this effect could be alleviated with increasing periods of continuous planting.

The immediate trends in atrial electrical remodeling for paroxysmal atrial fibrillation across different modes of catheter ablation.

BACKGROUND: Catheter ablation has emerged as a major strategy for paroxysmal atrial fibrillation (PAF). Atrial electrical remodeling (AER) plays a critical role in the recurrence of PAF after ablation. HYPOTHESIS: To characterize the immediate trends of AER during ablations in patients with PAF, and assess the relationship between immediate trends and recurrence. METHODS: We performed this prospective observational study of 135 patients to investigate AER following three ablation modes: radiofrequency ablation (RFA), cryoablation (CA) and 3D mapping-guided cryoablation (3D-CA). The atrial effective refractory period (AERP) and atrial conduction time (ACT) were measured via electrophysiology before and immediately after ablation, and P-wave indices were measured via electrocardiography before and within 24 h after ablation. Follow-up visits were conducted for at least 1 year or until relapse. RESULTS: Different approaches of ablation caused a fairly significant increase in the shortest P-wave duration and AERP in both the proximal coronary sinus (PCS) and distal coronary sinus (DCS) but caused a shortened P-wave dispersion. No different effect was found at the AERP among the three modes. Compared to patients who received CA, among patients who received RFA, a significant reduction in total ACT and right ACT was seen. Statistically, there was a weakly positive association between changes in total ACT and early recurrence. CONCLUSIONS: Injury during ablation for PAF was associated with an increase in the AERP but not in the ACT. Total ACT and right ACT were shorter after RFA than after CA. The increase in total ACT were slightly predictive of early recurrence.

Voltine Ecotypes of the Asian Corn Borer and Their Response to Climate Warming.

In the Asian corn borer (ACB), Ostrinia furnacalis (Guenée), diapause is governed by a multigenetic constitution that responds to daylength and temperature with seasonality. The ACB displays uni- or multivoltinism, depending on its geographic specificity. Hence, warmer temperatures may result in alternation of voltinism in the ACB, which will help in understanding the ecological consequences of climate warming on insects. In the present study, we investigated the voltinism in two natural populations from Harbin (H) and Gongzhuling (G) as well as a laboratory (L) population (established from the H population in 2017) of the ACB, at ambient and elevated atmospheric CO2 (aCO2 390 µL/L and eCO2 750 µL/L) and temperature (aT and Et = At + 2 °C). From the diapause response, both the uni- and multivoltine ecotypes were coexisting in the H population. The neonate occurrence date of 50% individuals that induced diapause was ca. 10 days later in the G population than in the H population, but it was about 10 days earlier than in the L population. Comparing to the dates of onset and the peak of diapause induction, the G and L populations were less variable than the H population in response to a short and/or shortening daylength in the field. The univoltine individuals could not be eliminated completely after 19 generations of selection. Diapause incidence decreased with a climate-warming scenario, which was temporally specific and could be overridden by significantly low daily average temperatures. The eCO2 did not directly impact the voltinism. On the basis of voltinism, the H population was sympatric for uni- and multivoltine ecotypes, with multivoltinism being dominant. The univoltinism trait was recessive. Climate warming could significantly override the effect of photoperiod, which was yearly dependent. Warmer temperatures and a decreased latitude (shortened daylength), and their interaction, would drive ACB evolution toward diapause homogeneity for multivoltinism.

Stimulated production of steroids in Inonotus obliquus by host factors from birch.

Steroids was considered as one of the bioactive components in Inonotus obliquus, while this kind of secondary metabolites are less accumulated in cultured mycelia. In this study, effect of extracts from bark and core of host-related species, birch (Betula platyphylla Suk.), on steroid production of I. obliquus in submerged culture were evaluated. The results showed that all dosages (0.01 and 0.1 g/L) of aqueous extracts and methanol extracts from birch bark and birch core possessed significantly stimulatory effect on steroid production of I. obliquus (P < 0.05). Among the eight extracts, the aqueous extract (0.01 g/L) from birch bark gave the highest steroid production (225.5 ± 8.7 mg/L), which is 97.3% higher than that of the control group. The aqueous extract (0.01 and 0.1 g/L) from birch bark could simultaneously stimulated mycelial growth and steroid content, while the methanol extract from birch bark only elevated the steroid content. High performance liquid chromatography analysis showed that productions of betulin, ergosterol, cholesterol, lanosterol, stigmasterol, and sitosterol in I. obliquus simultaneously increased in the presence of aqueous extract and methanol extract from birch bark. The results presented herein indicate that extracts from birch bark could act as an inducer for steroid biosynthesis of I. obliquus.