Mixed plantations do not necessarily provide higher ecosystem multifunctionality than monoculture plantations.

Forest stand transformation is a crucial strategy for enhancing the productivity and stability of planted forest ecosystems and maximizing their ecosystem functions. However, understanding forest ecosystem multifunctionality responses to various stand transformation methods remains limited. In this study, we assessed ecosystem multifunctionality, encompassing nutrient cycling, carbon stocks, water regulation, decomposition, wood production, and symbiosis, under different stand transformation methods (Chinese fir monoculture, mixed conifer and broad-leaf, broad-leaf mixed, and secondary forests). We also identified key factors contributing to variations in ecosystem multifunctionality. The results showed that Chinese fir plantations were more conducive to carbon stock creation, while broad-leaved mixed plantations excelled in water regulation. Secondary forests exhibited higher ecosystem multifunctionality than other plantation types, with Chinese fir plantations displaying the highest multifunctionality, significantly surpassing mixed coniferous and broad-leaved plantations. Our findings further revealed that soil nutrients and plant diversity have significant impacts on ecosystem multifunctionality. In summary, stand transformation profoundly influences ecosystem multifunctionality, and mixed plantations do not necessarily provide higher ecosystem multifunctionality than monoculture plantations.

Host-Parasitoid Phenology, Distribution, and Biological Control under Climate Change.

Climate change raises a serious threat to global entomofauna-the foundation of many ecosystems-by threatening species preservation and the ecosystem services they provide. Already, changes in climate-warming-are causing (i) sharp phenological mismatches among host-parasitoid systems by reducing the window of host susceptibility, leading to early emergence of either the host or its associated parasitoid and affecting mismatched species' fitness and abundance; (ii) shifting arthropods' expansion range towards higher altitudes, and therefore migratory pest infestations are more likely; and (iii) reducing biological control effectiveness by natural enemies, leading to potential pest outbreaks. Here, we provided an overview of the warming consequences on biodiversity and functionality of agroecosystems, highlighting the vital role that phenology plays in ecology. Also, we discussed how phenological mismatches would affect biological control efficacy, since an accurate description of stage differentiation (metamorphosis) of a pest and its associated natural enemy is crucial in order to know the exact time of the host susceptibility/suitability or stage when the parasitoids are able to optimize their parasitization or performance. Campaigns regarding landscape structure/heterogeneity, reduction of pesticides, and modelling approaches are urgently needed in order to safeguard populations of natural enemies in a future warmer world.

Combined Exogenous Activation of Bovine Oocytes: Effects on Maturation-Promoting Factor, Mitogen-Activated Protein Kinases, and Embryonic Competence.

Oocyte activation via dual inhibition of protein synthesis and phosphorylation has improved in vitro embryo production in different mammalian species. In this study, we evaluated the effects of the combination of cycloheximide (CHX), dimethyl amino purine (DMAP), and anisomycin (ANY) on the activation of bovine oocytes, particularly on dynamics of MPF and MAPKs, embryonic developmental potential, and quality. The results showed that the cleavage and blastocyst rates, as well as levels of CCNB1, CDK1, p-CDK1Thr161, and p-CDK1Thr14-Tyr15, were similar among groups; ANY and ANY + CHX reduced the expression of ERK1/2 compared to DMAP-combinations (p < 0.05), whereas ANY + DMAP, CHX + DMAP, and ANY + CHX + DMAP reduced p-ERK1/2 compared to ANY and ANY + CHX treatments (p < 0.05). The quality of blastocysts in terms of cell counts, their allocation, and the numbers of TUNEL-positive cells did not differ among groups. However, transcript levels of POU5F1 were higher in embryos derived from ANY + CHX + DMAP treatment compared to other groups, while expression levels of CDX2 did not show differences. In addition, the BCL2A1/BAX ratio of the ANY + CHX + DMAP treatment was significantly low compared to the ANY treatment (p < 0.05) and did not differ significantly from the other treatments. In conclusion, oocyte activation by dual inhibition of protein synthesis and phosphorylation induces MPF inactivation without degradation of CCNB1, while MAPK inactivation occurs differentially between these inhibitors. Thus, although the combined use of these inhibitors does not affect early developmental competence in vitro, it positively impacts the expression of transcripts associated with embryonic quality.

Haploid androgenetic development of bovine embryos reveals imbalanced WNT signaling and impaired cell fate differentiation†.

Haploid embryos have contributed significantly to our understanding of the role of parental genomes in development and can be applied to important biotechnology for human and animal species. However, development to the blastocyst stage is severely hindered in bovine haploid androgenetic embryos (hAE). To further our understanding of such developmental arrest, we performed a comprehensive comparison of the transcriptomic profile of morula-stage embryos, which were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) of transcripts associated with differentiation in haploid and biparental embryos. Among numerous disturbances, results showed that pluripotency pathways, especially the wingless-related integration site (WNT) signaling, were particularly unbalanced in hAE. Moreover, transcript levels of KLF4, NANOG, POU5F1, SOX2, CDX2, CTNNBL1, AXIN2, and GSK3B were noticeably altered in hAE, suggesting disturbance of pluripotency and canonical WNT pathways. To evaluate the role of WNT on hAE competence, we exposed early Day-5 morula stage embryos to the GSK3B inhibitor CHIR99021. Although no alterations were observed in pluripotency and WNT-related transcripts, exposure to CHIR99021 improved their ability to reach the blastocysts stage, confirming the importance of the WNT pathway in the developmental outcome of bovine hAE.

Carbon sequestration and storage capacity of Chinese fir at different stand ages.

In southern China, Chinese fir Cunninghamia lanceolata is one of the most important native conifer trees, widely used in afforestation programs. This area has the largest forestland atmospheric carbon sink, and a relatively young stand age characterizes these forests. However, how C. lanceolata forests evolved regarding their ability to sequester carbon remains unclear. Here we present data on carbon storage and sequestration capacity of C. lanceolata at six stand ages (5-, 10-, 15-, 20-, 30- and 60 - year-old stands). Results show that the carbon stock in trees, understory, vegetation, litter, soil, and ecosystem significantly increased with forest age. The total ecosystem carbon stock increased from 129.11 to 348.43 Mg ha-1 in the 5- and 60 - year-old stands. The carbon sequestration rate of C. lanceolata shows an overall increase in the first two stand intervals (5-10 and 10-15), peaks in the 15-20 stand intervals, and then decreases in the 20-30 and 30-60 stand intervals. Our result revealed that carbon sequestration rate is a matter of tree age, with the highest sequestration rates occurring in the middle age forest (15-20 - year-old). Therefore, this information may be useful for national climate change mitigation actions and afforestation programs, since forests are primarily planted for this purpose.

Standardization of a Sex-Sorting Protocol for Stallion Spermatozoa by Means of Absolute RT-qPCR.

Sperm sexing is a technology that can generate great economic benefits in the animal production sector. Techniques such as sex-sorting promise over 90% accuracy in sperm sexing. However, for the correct standardization of the technique, some laboratory methodologies are required. The present manuscript describes in detail a standardized equine sperm sex-sorting protocol using an absolute qPCR-based methodology. Furthermore, the results of absolute qPCR were implemented and validated by generating equine/bovine heterologous embryos by intracytoplasmic sperm injection (ICSI) of presumably sexed equine spermatozoa into bovine oocytes using a piezoelectric system (Piezo-ICSI). Our results indicated that equine sex-sorting spermatozoa had a 97% and 94% certainty for X and Y sperm, respectively, while presumptive female and male equine/bovine hybrid embryos, generated by Piezo-ICSI, had an accuracy of 92% with respect to the desired sex. Therefore, it is concluded that the presented methodology is a reliable, cost-effective, and relatively simple option for standardizing sex-sorting of equine spermatozoa. This is supported by the results of the correct sexing of Piezo-ICSI heterologous embryos generated with the sexed spermatozoa, validating the correct sexing and viability of these gametes.

Effect of Atomized Black Maca (Lepidium meyenii) Supplementation in the Cryopreservation of Alpaca (Vicugna pacos) Epididymal Spermatozoa.

Artificial insemination is an important assisted reproductive technology that has been applied in several mammalian species. However, successful cryopreservation of semen of South American camelids has been limited, hindering the commercial application of artificial insemination in alpaca species. In this scenario, the addition of antioxidants to semen extenders provides a strategy to improve the freezability of mammalian sperm. Bioactive metabolites from natural extracts of black maca have shown valuable antioxidant properties. Thus, the objective of this study was to evaluate the effect of the addition of atomized black maca in the freezing medium of epididymal spermatozoa of alpacas. Fifteen pairs of epididymis were collected from a local slaughterhouse. Each sample was divided into six groups: (1) fresh, (2) yolk medium (YM), (3) 10 mg/mL maca, (4) 20 mg/mL maca, (5) 30 mg/mL maca, and (6) resveratrol (as an antioxidant control). Sperm cryopreservation was performed through the slow freezing method. Markers associated with functionality, such as motility, viability, and plasma membrane integrity, as well as markers associated with oxidative damage, such as DNA integrity, total ROS production, and mitochondrial function, were analyzed. The results show that the supplementation with black maca (20 mg/mL) improved the sperm motility, viability, plasma membrane integrity, and mitochondrial function evaluated according to an index of formazan deposits. Similarly, the ROS production decreased with maca at 20 mg/mL, although the DNA integrity did not show any differences among the groups. These results suggest that maca at 20 mg/mL has cytoprotective effects during freezing/thawing of epididymal sperm of alpaca species. Further research will be focused on assessing the effects of maca supplementation on semen extenders by using biomolecular markers (proAKAP4) associated with fertility.

Acidity of Soil and Water Decreases in Acid-Sensitive Forests of Tropical China.

Acid deposition in China has been declining since the 2000s. While this may help mitigate acidification in forest soils and water, little is known about the recovery of soils and water from previous severe acidification in tropical China. Here, we assessed the chemistry of mineral soils, water, and acid gases (SO2 and NOx) from three successional forest types in tropical China from 2000 to 2022. Our results showed that soil pH increased synchronously from 3.9 (2000-2015) to 4.2 (2016-2022) across all three forest types, with exchangeable acid initially decreasing and thereafter stabilizing. Surface and ground water pH also gradually increased throughout the monitoring period. Soil pH recovery was stronger in the primary than in the planted forest. However, soil pH recovery lagged behind the increase in rainfall pH by approximately a decade. The recovery of soil pH was likely related to the positive effects of the dissolution of Al/Fe-hydroxysulfate mineral and subsequent sulfur desorption on soil acid-neutralizing capacity, increased soil organic matter, and climate warming, but was likely moderated by increased exchangeable aluminum and potentially proton-producing hydroxysulfate mineral dissolution that caused the lagged soil pH recovery. Surface and ground water pH recovery was attributed to increased water acid-neutralizing capacity. Our study reports the potential for the recovery of acidified soil and water following decreased acid deposition and provides new insights into the functional recovery of acid-sensitive forests.

Carbon storage capacity of Castanopsis hystrix plantations at different stand-ages in South China.

The establishment of forest plantations is an excellent silvicultural alternative to improve atmospheric carbon (C) sequestration for climate change mitigation. In recent years, the importance of forest growth and C dynamics at different stand ages have drawn huge attention for investigation. To evaluate the impacts of different stand ages on the C storage capacity of Castanopsis hystrix plantations ecosystems, we measured biomass and C content in trees, understory, litter, and soil components from field measurements at different stand ages, i.e., 6-, 10-, 15-, 25-, and 34-year-old stands. Results showed that the C stocks in trees, understory vegetation, litter, soil and the whole ecosystem increased steadily with stand age. C. hystrix C storage capacity in the 6-, 10-, 15-, 25-, and 34-year-old stands were 29.17, 56.18, 85.60, 132.19 and 157.79 Mg ha-1, respectively, while the total ecosystem C storage capacity was 68.11, 106.11, 136.13, 187.34 and 226.14 Mg ha-1, respectively. Meanwhile, the C sequestration rate in the 6-10, 10-15, 15-25 and 25-35-year-old stands were 675.28, 588.35, 465.97 and 284.39 g m2 year-1, respectively. Similarly, total biomass of trees, understory, and litter increased with stand age. Our study indicated that C. hystrix plantations are still developing in the area, since live biomass and soil C continue to accumulate despite reduced C sequestration rates. The information provided here highlight the C stock and C sequestration rates of C. hystrix at different stand ages, providing baseline information on the C dynamics of young and older stand of C. hystrix in this region; Hence, such knowledge could be useful for designing more realistic policies to mitigate climate change, such as programs to maintain continuous forest growth.

Genome-wide identification and immune response analysis of mitogen-activated protein kinase cascades in tea geometrid, Ectropis grisescens Warren (Geometridae, Lepidoptera).

BACKGROUND: Tea geometrid Ectropis grisescens (Geometridae: Lepidoptera), is one of the most destructive defoliators in tea plantations in China. The MAPK cascade is known to be an evolutionarily conserved signaling module, acting as pivotal cores of host-pathogen interactions. Although the chromosome-level reference genome of E. grisescens was published, the whole MAPK cascade gene family has not been fully identified yet, especially the expression patterns of MAPK cascade gene family members upon an ecological biopesticide, Metarhizium anisopliae, remains to be understood. RESULTS: In this study, we have identified 19 MAPK cascade gene family members in E. grisescens, including 5 MAPKs, 4 MAP2Ks, 8 MAP3Ks, and 2 MAP4Ks. The molecular evolution characteristics of the whole Eg-MAPK cascade gene family, including gene structures, protein structural organization, chromosomal localization, orthologs construction and gene duplication, were systematically investigated. Our results showed that the members of Eg-MAPK cascade gene family were unevenly distributed in 13 chromosomes, and the clustered members in each group shared similar structures of the genes and proteins. Gene expression data revealed that MAPK cascade genes were expressed in all four developmental stages of E. grisescens and were fairly and evenly distributed in four different larva tissues. Importantly, most of the MAPK cascade genes were induced or constitutively expressed upon M. anisopliae infection. CONCLUSIONS: In summary, the present study was one of few studies on MAPK cascade gene in E. grisescens. The characterization and expression profiles of Eg-MAPK cascades genes might help develop new ecofriendly biological insecticides to protect tea trees.

Baseline Analysis of Endophytic Fungal Associates of Solenopsis invicta Buren from Mounds across Five Counties of Guangdong Province, China.

Red imported fire ants mounds have been suggested as a potential reservoir for beneficial entomopathogenic fungal species that are vital for more complex roles in the ecosystem aside from infecting the insects. In the current study, the assemblage of fungal symbionts of the red imported fire ants (RIFA) were obtained across five cities in Guangdong Province, China. The sampling areas were selected because of high occurrence of fire ants mounds in the regions. Mound soils, plant debris within mounds, and ants were collected from three sampling locations in each city for potential isolation of entomopathogenic fungal associates of RIFA. All samples were collected during the spring of 2021. Following successful isolation from substrates, the patterns of fungal species composition, and richness were evaluated. In total, 843 isolates were recovered, and based on their phenotypic distinctiveness and molecular characterization based on DNA sequences of multiple loci including the ITS, SSU, and LSU regions, 46 fungal taxa were obtained, including 12 that were unidentified. Species richness and abundance was highest in the mound soils, while the lowest value was recorded from the ant body. As per the different locations, the highest abundance level was recorded in Zhuhai, where 15 fungal taxa were cultivated. The most common taxa across all substrates and locations was Talaromyces diversus. A baseline analysis of the fungal community composition of RIFA would better our understanding on the interactions between these social ants and their associated microbial organisms, and this knowledge in turn would be important for the successful management of the RIFA.

Comprehensive genome-wide identification and expression profiling of ADF gene family in Citrus sinensis, induced by endophytic colonization of Beauveria bassiana.

Endophytic entomopathogenic species are known to systematically colonize host plants and form symbiotic associations that benefit the plants they live with. The actin-depolymerizing factors (ADFs) are a group of gene family that regulate growth, development, and defense-related functions in plants. Systematic studies of ADF family at the genome-wide level and their expression in response to endophytic colonization are essential to understand its functions but are currently lacking in this field. 14ADF genes were identified and characterized in the Citrus sinensis genome. The ADF genes of C. sinensis were classified into five groups according to the phylogenetic analysis of plant ADFs. Additionally, the cis-acting analysis revealed that these genes play essential role in plant growth/development, phytohormone, and biotic and abiotic responses; and the expression analysis showed that the symbiotic interactions generate a significant expression regulation level of ADF genes in leaves, stems and roots, compared to controls; thus enhancing seedlings' growth. Additionally, the 3D structures of the ADF domain were highly conserved during evolution. These results will be helpful for further functional validation of ADFs candidate genes and provide important insights into the vegetative growth, development and stress tolerance of C. sinensis in responses to endophytic colonization by B. bassiana.

Simple and Efficient Chemically Defined In Vitro Maturation and Embryo Culture System for Bovine Embryos.

Supplementation of the culture media for in vitro production (IVP) of bovine embryos with fetal bovine serum (FBS) is associated with inconsistent outcomes. The present study sought to replace FBS and BSA by insulin-like growth factor 1 (IGF1), fibroblast growth factor 2 (FGF2) and epidermal growth factor (EGF). In Experiment 1, absence of FBS from maturation medium (MM) did not affect the rate of in vitro maturation, as assessed by the extrusion of the first polar body. However, when gonadotropins and FBS were removed from the MM, the maturation rate was significantly reduced even in the presence of growth factors. Therefore, gonadotropin-supplemented MM medium was established as the base medium for the defined maturation condition. In Experiment 2, the addition of growth factors to gonadotropin-supplemented MM medium supported similar maturation (~90%) compared to the undefined condition (FBS-carrying). In Experiment 3, the addition of growth factors to embryo culture medium showed similar in vitro competence compared to the undefined (FBS) control. In Experiment 4, completely defined conditions (absence of FBS and BSA during in vitro maturation and embryo culture) were tested. A higher cleavage was observed with FGF2 (86%) compared to EGF (77%) and the FBS control (77%), but similar blastocyst rates were observed for FGF2 (24%), EGF (19%) and the FBS control (25%). Embryo quality was similar among groups. Finally, post-thawing survival was higher for FGF2 (94%) compared to the FBS control (77%). Thus, we report a simple defined IVP system for bovine species that generates developmental outcomes and embryos of similar quality than those produced under conditions containing FBS.

Genome-Wide Identification and Characterization of Toll-like Receptors (TLRs) in Diaphorina citri and Their Expression Patterns Induced by the Endophyte Beauveria bassiana.

Toll-like receptors (TLRs) are pathogen recognition receptors (PRRs), which play key roles in helping the host immune system fight pathogen invasions. Systematic information on TLRs at the genome-wide level and expression profiling in response to endophytic colonization is very important to understand their functions but is currently lacking in this field. Here, a total of two TLR genes were identified and characterized in Diaphorina citri. The TLR genes of D. citri were clustered into five families according to the phylogenetic analysis of different species' TLRs. The domain organization analyses suggested that the TLRs were constituted of three important parts: a leucine-rich repeat (LRR) domain, a transmembrane region (TR) and a Toll/interleukin-1 receptor (TIR) domain. The mRNA expression levels of the two TLR genes (DcTOLL and DcTLR7) were highly regulated in both nymphs and adults of D. citri. These results elucidated the potentiated TLR gene expression in response to endophytically colonized plants. Furthermore, the 3D structures of the TIR domain were highly conserved during evolution. Collectively, these findings elucidate the crucial roles of TLRs in the immune response of D. citri to entomopathogens systematically established as endophytes, and provide fundamental knowledge for further understanding of the innate immunity of D. citri.

High prevalence and clinical characteristics of respiratory infection by human rhinovirus in children from Lima-Peru during years 2009-2010.

INTRODUCTION: Human rhinovirus is a major cause of acute respiratory infections (ARIs) worldwide. Epidemiological data on human rhinovirus (RV) in Peru is still scarce, as well as its role in respiratory infections in children. Therefore, the aim of this study was to describe the prevalence of rhinovirus and to identify the circulating species in nasopharyngeal swabs from children with acute respiratory infections. MATERIALS AND METHODS: We analyzed nasopharyngeal swab samples that were collected from children younger than 17 years old, who had a clinical diagnosis of ARI from the "Hospital Nacional Cayetano Heredia" between May 2009 and December 2010. The original study recruited 767 inpatients with ARI, 559 samples of which were included and analyzed in the current study. Detection of rhinovirus and determination of rhinovirus species were characterized by PCR. RESULTS: Rhinovirus was detected in 42.22% samples (236/559), RV-A was detected in 10.17% (24/236) of the cases, RV-B in 16.53% (39/236), and RV-C in 73.31% (173/236). The age group with the highest number of cases was the 0-5 months group with 45.97%, followed by the 1-5 years group with 25.22%. Most of the positive RV cases, i.e., 86.44% (204/236), were hospitalized. The most common signs and symptoms found in patients who tested positive for RV were cough (72.88%), fever (68.64%), rhinorrhea (68.22%), and respiratory distress (61.44%). Infection with RV-A was associated with wheezing (p = 0.02). Furthermore, RV-C was related to cough (p = 0.01), wheezing (p = 0.002), and conjunctival injection (p = 0.03). A peak in RV-C cases was found in March (32 cases in 2010); June (18 cases in 2009 and 12 cases in 2010), which corresponds to the fall season in Peru; and also November (17 cases in 2009 and 4 cases in 2010), which corresponds to spring. RV-A and RV-B cases were constant throughout the year. CONCLUSION: In conclusion, we found a high prevalence of rhinovirus C infection among pediatric patients with acute respiratory infections in Lima, Peru. This viral infection was more common in children between 0 to 5 months old, and was associated with cough, wheezing, and conjunctival injection. Epidemiological surveillance of this virus should be strengthened/encouraged in Peru to determine its real impact on respiratory infections.

Pluripotent Core in Bovine Embryos: A Review.

Early development in mammals is characterized by the ability of each cell to produce a complete organism plus the extraembryonic, or placental, cells, defined as pluripotency. During subsequent development, pluripotency is lost, and cells begin to differentiate to a particular cell fate. This review summarizes the current knowledge of pluripotency features of bovine embryos cultured in vitro, focusing on the core of pluripotency genes (OCT4, NANOG, SOX2, and CDX2), and main chemical strategies for controlling pluripotent networks during early development. Finally, we discuss the applicability of manipulating pluripotency during the morula to blastocyst transition in cattle species.

Comparative microbiome analysis of Diaphorina citri and its associated parasitoids Tamarixia radiata and Diaphorencyrtus aligarhensis reveals Wolbachia as a dominant endosymbiont.

Microbiome analysis in a host-parasitoid interaction network was conducted to compare the taxonomic composition of bacterial communities of Diaphornia citri, Tamarixia radiata, and Diaphorencyrtus aligarhensis. The comparative analysis revealed differences in the composition and diversity of the symbiont populations across the host and its associated parasitoids. Proteobacteria was the most dominant phylum, representing 67.80% of the total bacterial community, while Candidatus Profftella armature and Wolbachia were the dominant genera across the host and parasitoids. There were clear differences observed in alpha and beta diversity of microbiota through the host and its associated parasitoids. The function prediction of bacterial communities and Pearson correlation analysis showed that specific bacterial communities displayed positive correlations with the carbohydrate metabolism pathway. Furthermore, when symbiotic bacteria were eliminated using a broad-spectrum antibiotic, tetracycline hydrochloride, the parasitoids' median survival time and longevity were significantly reduced. We confirmed the physiological effects of symbiotic bacteria on the fitness of parasitoids and demonstrated the effect of antibiotics in decreasing the food intake and measurement of amino acids in the hemolymph. This study sheds light on basic information about the mutualism between parasitoids and bacteria, which may be a potential source for biocontrol strategies for citrus psyllid, especially D. citri.

Comparative transcriptome analysis of Tamarixia radiata (Hymenoptera: Eulophidae) reveals differentially expressed genes upon heat shock.

Temperature is a key parameter that affects insect population, abundance, and distribution in tropical and subtropical regions. Tamarixia radiata Waterson (Hymenoptera: Eulophidae) is a species-specific ectoparasitoid widely used as a biological control agent for the major citrus pest Diphornia citri Kuwayama (Hemiptera: Liviidea). To date, T. radiata response to high temperature at the molecular level still is unclear. In this study, we conducted a comparative analysis of the transcriptomes of T. radiata exposed at 25 °C and 38 °C for 15 min. A total of 51,072 unigenes were obtained, 22,413 annotated with a mean length of 1054 bp. Differential expression analysis showed that 502 genes were identified, including 476 genes significantly up-regulated and 26 genes down-regulated after heat stress exposure. The Gene Ontology analysis showed that most enriched DEGs are categorized into "cellular process", "metabolic process" and "DNA binding." In addition, "Lysosome," "Longevity regulating pathway-multiple species," and "starch and sucrose metabolism" were highly enriched in Kyoto Encyclopedia of Genes and Genomes pathways. Transcriptome analyses showed that heat stress significantly induced the transcription of the molecular chaperone, immune response, stress signaling transduction, and oxidation resistance, including highly expressed heat shock proteins, ATPases, and detoxifying enzymes. Furthermore, the expression patterns of thirteen genes including heat shock proteins (HSP), glutathione S-transferase (GST) and cytochrome P450 were consistent with the transcriptome results obtained through qRT-PCR. Together, our results provided a comprehensive study of the molecular response of T. radiata to heat stress and provides new insight for the future functional validation of heat resistance-related genes.

General Limitations to Endophytic Entomopathogenic Fungi Use as Plant Growth Promoters, Pests and Pathogens Biocontrol Agents.

The multiple roles of fungal entomopathogens in host plants' growth promotion, pest and pathogen management have drawn huge attention for investigation. Endophytic species are known to influence various activities of their associated host plants, and the endophyte-colonized plants have been demonstrated to gain huge benefits from these symbiotic associations. The potential application of fungal endophytes as alternative to inorganic fertilizers for crop improvement has often been proposed. Similarly, various strains of insect pathogenic fungi have been formulated for use as mycopesticides and have been suggested as long-term replacement for the synthetic pesticides that are commonly in use. The numerous concerns about the negative effects of synthetic chemical pesticides have also driven attention towards developing eco-friendly pest management techniques. However, several factors have been underlined to be militating the successful adoption of entomopathogenic fungi and fungal endophytes as plant promoting, pests and diseases control bio-agents. The difficulties in isolation and characterization of novel strains, negative effects of geographical location, vegetation type and human disturbance on fungal entomopathogens, are among the numerous setbacks that have been documented. Although, the latest advances in biotechnology and microbial studies have provided means of overcoming many of these problems. For instance, studies have suggested measures for mitigating the negative effects of biotic and abiotic stressors on entomopathogenic fungi in inundative application on the field, or when applied in the form of fungal endophytes. In spite of these efforts, more studies are needed to be done to achieve the goal of improving the overall effectiveness and increase in the level of acceptance of entomopathogenic fungi and their products as an integral part of the integrated pest management programs, as well as potential adoption as an alternative to inorganic fertilizers and pesticides.