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Tinospora sagittata is a columbin-rich medicinal plant, but its columbin levels are reduced under artificial cultivation conditions. The objective of this study was to analyze the effects of inoculations with Diversispora versiformis (Dv), Funneliformis mosseae (Fm), Rhizophagus intraradices (Ri), and mixed inoculation (Dv + Fm + Ri) (Mix) on growth performance, root morphology, leaf photosynthetic physiology, and root columbin levels in T. sagittata. These arbuscular mycorrhizal fungi (AMF) were able to colonize the roots, as evidenced by a root mycorrhizal colonization rate ranging from 17% to 48% and soil hyphal lengths ranging from 17.51 cm/g to 32.02 cm/g, with the Mix treatment being the greatest. AMF inoculations improved plant height (16–151%), leaf number (119–283%), shoot (37–211%), and root biomass (22–318%) to varying extents, with Ri and Mix treatments being the most prominent. AMF-treated plants presented relatively greater root total length, projected area, surface area, volume, and average diameter, especially those treated with Ri and Mix. AMF inoculations also significantly improved the leaf nitrogen balance index, transpiration rate, and stomatal conductance, while the photosynthesis rate and chlorophyll index varied by AMF species, along with a decrease in intercellular CO2 levels. Root columbin levels ranged from 0.524 mg/g to 5.389 mg/g, and AMF inoculation significantly increased root columbin levels by 228–928%, with Ri being the most significant. Root columbin levels were significantly positively correlated with soil hyphal length, root total length, root projected area, root surface area, root volume, and root average diameter, but not root AMF colonization rate. This study demonstrates for the first time that AMF, especially Ri, can be employed as a biostimulant to promote growth as well as root columbin levels in T. sagittata, where AMF-triggered improvement in root morphology is an important reason for promoting root columbin levels. [ABSTRACT FROM AUTHOR]

Either arbuscular mycorrhizal (AM) fungi or earthworms as soil organisms promote plant growth, while their interaction on nitrogen (N) content in plants, soil, and glomalin-related soil protein (GRSP) is not known. In this study, an AM fungus (Funneliformis mosseae) and an earthworm (Pheretima guillelmi), either in single or in combination, were introduced into soil growing white clover to analyze their effects on plant growth, chlorophyll, soluble protein, N contents in leaves, roots, GRSP, and soil, and the contribution of N in purified GRSP to soil total N. The four-week introduction of earthworms significantly increased root mycorrhizal colonization rate, and accelerated an improved AM effect on chlorophyll and plant growth. The single introduction of earthworms significantly increased root N and soil total N contents, while the single introduction of AM fungi distinctly elevated N contents in leaves, roots, easily extractable GRSP, and soil (nitrate N and total N). The N in total GRSP was 5.78–7.70 mg g−1, accounting for 3.46 %–5.45 % of the soil total N, of which the contribution of N from easily extractable GRSP and difficult-to-extract GRSP was 1.84 %–3.07 % and 1.34 %–2.39 %, respectively. AM fungi, but not earthworms, significantly increased the contribution of N in GRSP to soil total N, and the introduction of earthworms further accelerated the increased effect of mycorrhizas on the contribution of N in easily extractable and total GRSP to soil total N. These results demonstrated that the introduction of earthworms into mycorrhizosphere can facilitate N storage in GRSP and thus a contribution to soil total N. • Effects of earthworms and AMF on N content of soil and plants were studied. • Dual earthworms and AMF addition recorded higher plant growth and chlorophyll than single addition. • Earthworms increased root N and soil total N, while AMF elevated N in plants, EE-GRSP, and soil NO 3 −−N and total N. • Dual addition recorded higher N contents in GRSPs than single addition. • Earthworm accelerated the AMF effect on the contribution of N in EE-GRSP and T-GRSP to soil total N. [ABSTRACT FROM AUTHOR]

Earthworms and arbuscular mycorrhizal fungi (AMF) are beneficial organisms in the soil and play an important role in improving soil fertility and plant growth. The objective of this study was to analyze the response of single versus dual inoculation of earthworms (Pheretima guillelmi) and AMF (Funneliformis mosseae) on mycorrhizal growth, root morphology, and rhizosphere properties of white clover (Trifolium repens). Addition of earthworms significantly increased the degree of root AMF colonization, while decreased hyphal length and spore number in the soil. Single or dual inoculation of F. mosseae and P. guillelmi significantly increased root traits (total length, area, volume, and average diameter), soil phosphatase activities (acid, neutral, and alkaline), easily extractable and total glomalin-related soil protein content, the percentage of soil water-stable aggregates at the size of 2–4 mm, 1–2 mm and 0.5–1 mm, aggregate stability, Bray-P, and soil organic carbon (SOC) contents. Among them, the dual inoculation further amplified the effect on glomalin production, phosphatase activity, and aggregate stability, but did not show a superposition effect on root improvement, Bray-P, and SOC. Our study, hence, suggested that AMF and earthworms have synergistic roles in elevating rhizosphere properties vis-à-vis agronomic responses of white clover. • Effects of earthworm-AMF (singly and in combination) on rhizosphere environment and crop response were studied. • Earthworms increased root AMF colonization, but reduced hyphae and spores in soils. • AMF or earthworms singly improved root traits, Bray-P, and soil organic carbon contents, but dual application of AMF and earthworms did not show a major effect than single application. • Single AMF or earthworms increased aggregation, phosphatase, GRSP, and dual addition amplified these responses. [ABSTRACT FROM AUTHOR]

Endophytes have the ability to improve plant nutrition alongside their agronomic performance, among which arbuscular mycorrhizal fungi provide the most benefits to their host. Previously, we reported for the first time that an arbuscular mycorrhizal-like fungus Piriformospora indica had the ability to colonize roots of trifoliate orange ( Poncirus trifoliata ) and conferred positive effects on nutrient acquisition. Present study showed the changes in fatty acids and sugars to unravel the physiological and symbiotic association of trifoliate orange with P . indica and an arbuscular mycorrhizal fungus, Funneliformis mosseae singly or in combination. All the endophytic fungi collectively increased fructose, glucose, and sucrose content in leaves and roots, along with a relatively higher increase with P . indica inoculation than with F . mosseae alone or dual inoculation. Treatment with P . indica increased the concentration of part unsaturated fatty acids such as C18:3N6, C20:2, C20:3N6, C20:4N6, C20:3N3, C20:5N3, C22:1N9, and C24:1. Additionally, P . indica induced the increase in the concentration of part saturated fatty acids such as C6:0, C8:0, C13:0, C14:0, and C24:0. F . mosseae hardly changed the content of fatty acids, except for increase in C14:0 and C20:5N3. Double inoculation only reduced the C21:0, C10:0, C12:0, C18:3N3, and C18:1 content and increased the C20:5N3 content. These endophytic fungi up-regulated the root PtFAD2, PtFAD6, PtΔ9 , and PtΔ15 gene expression level, coupled with a higher expression of PtFAD2 and PtΔ9 by P . indica than by F . mosseae . It was concluded that P . indica exhibited a stronger response, for sugars and fatty acids, than F . mosseae on trifoliate orange. Such results also reveal the Pi (an in vitro culturable fungus) as a bio-stimulator applying to citriculture.

Dubin-Johnson syndrome (DJS) is an autosomal recessive disorder resulting from biallelic mutations of ABCC2 gene, with long-term or intermittent conjugated hyperbilirubinemia being the main clinical manifestation. This paper aims to report the clinical features and ABCC2 genotypes of an infant with DJS. A 9.5-month-old male infant was referred to the hospital due to abnormal liver function discovered over 9 months. The major clinical presentation was prolonged jaundice since neonatal period. A series of biochemistry analysis revealed markedly elevated total bilirubin, conjugated bilirubin and total bile acids. The patient had been managed in different hospitals, but the therapeutic effects were unsatisfactory due to undetermined etiology. Physical examination revealed jaundiced skin and sclera, and a palpable liver 3 cm below the right subcostal margin with medium texture. The spleen was not enlarged. Genetic analysis revealed a splice-site variant c.3988-2A>T and a nonsense variant c.3825C>G (p.Y1275X) in the ABCC2 gene of the infant, which were inherited from his mother and father respectively. The former had not been previously reported. Then ursodeoxycholic acid and phenobarbital were given orally. Half a month later, as a result, his jaundice disappeared and the biochemistry indices improved. However, the long-term outcome needs to be observed. Literature review revealed that neonates/infants with DJS presented with cholestatic jaundice soon after birth as the major clinical feature, and the ABCC2 variants exhibited marked heterogeneity.

Objective: To detect potential mutations of OTC gene in a male infant affected with ornithine transcarbamylase deficiency., Methods: Genomic DNA were isolated from peripheral blood samples of family members and 100 healthy individuals. Potential mutations of the 10 exons of OTC gene were screened with PCR and Sanger sequencing., Results: A homozygous missense mutation c.917G>C in exon 9, which results in p.R306T, was identified in the infant. Sequencing of the mother and two female members of the family indicated a heterozygous status for the same mutation. The same mutation was not found in other members of the family and 100 healthy controls., Conclusion: A missense mutation c.917G>C in the OTC gene is responsible for the pathogenesis of the disease. Identification of the mutation can facilitate prenatal diagnosis and genetic counseling for the family.

VH and Vkappa genes were amplified from human ScFv B17 specific against botulinum neurotoxin serotype A (BoNTa). Gene sequence encoding 5'-terminal 12 amino acid of heavy chain constant region CH1, as a linker, linked VH and Vkappa to construct a new three-domain antibody molecule VH/Vkappa. VH/Vkappa was expressed at high level over 34% of total host cell proteins in E. coli. Recombinant protein were purified up to 95% by affinity column. As a result, recombinant VH/Vkappa could recognize and bind specific to BoNTa in ELISA. However, comparing with its parent ScFv, VH/Vkappa has higher relative affinity and stability.