Hongming Tan

Isolation and characterization of endophytic Streptomyces strains from surface-sterilized tomato (Lycopersicon esculentum) roots

Aims:  To isolate endophytic Streptomyces strains from tomato and examine their antimicrobial activity.

Isolation of Endophytic Actinomycetes From Roots and Leaves of Banana (Musa Acuminata) Plants and Their Activities Against Fusarium oxysporumf. sp. cubense

Two hundred and forty-two actinomycete strains were isolated from the interior of leaves and roots of healthy and wilting banana plants. Most of them were streptomycetes, Streptomyces griseorubiginosus-like strains were the most frequently isolated strains. Community analysis demonstrated increased actinomycete diversity in wilting leaves compared to that in healthy leaves, similar actinomycete communities were found in wilting and healthy roots. Screening of the isolates for antagonistic activity against Fusarium oxysporumf. sp. cubenserevealed that the proportion of antagonistic streptomycetes in healthy roots was higher than that in wilting roots (P < 0.01), but no difference was found between antagonistic strains isolated from healthy and wilting leaves. The potential biological control of Panama disease of banana by endophytic streptomycetes, especially Streptomyces griseorubiginosus-like strains was discussed.

Characterization of a new keratinolytic Trichoderma atroviride strain F6 that completely degrades native chicken feather

Aims: To isolate novel nonpathogenic fungus that completely degrades native chicken feather and characterize its keratinases.

Isolation and characterization of actinomycetes from healthy goat faeces.

Aims:  To isolate and characterize actinomycetes with probiotic activities from healthy goat faeces.

Characterization of Heavy Metal-Resistant Endophytic Yeast Cryptococcus sp. CBSB78 from Rapes (Brassica chinensis) and Its Potential in Promoting the Growth of Brassica spp. in Metal-Contaminated Soils

A Cd-, Pb-, Zn-, Cu-resistant endophytic yeast CBSB78 was isolated from surface-sterilized rape roots. The isolate was identified as Cryptococcus sp. based on the ITS1–5.8S–ITS2 sequence analysis. The strain was resistant to 20 mM Cd2+, 20 mM Pb2+, 10 mM Zn2+, and 7 mM Cu2+. The yeast CBSB78 was a low indole-3-acetic acid (IAA) producer and possessed low 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Overall, 29.4–244 % of survival rates increased and the dry weight of Brassica alboglabra showed a 41.1 % increase when it was inoculated into the seedlings. The inoculation of CBSB78 could also increase the extraction amounts of Cd, Pb, and Zn by B. alboglabra simultaneously in the multi-metal contaminated soils, which showed the potential to improve extraction efficacy of Cd, Pb, Zn by B. alboglabra seedlings in the field.

Enhanced phytoremediation of toxic metals by inoculating endophytic Enterobacter sp. CBSB1 expressing bifunctional glutathione synthase.

To engineer plant-bacteria symbionts for remediating complex sites contaminated with multiple metals, the bifunctional glutathione (GSH) synthase gene gcsgs was introduced into endophytic Enterobacter sp. CBSB1 to improve phytoremediation efficiency of host plant Brassica juncea. The GSH contents of shoots inoculated with CBSB1 is 0.4μMg(-1) fresh weight. However, the GSH concentration of shoots with engineered CBSB1-GCSGS increased to 0.7μMg(-1) fresh weight. The shoot length, fresh weight and dry weight of seedlings inoculated with CBSB1-GCSGS increased 67%, 123%, and 160%, compared with seedlings without inoculation, respectively. The Cd and Pb concentration in shoots with CBSB1-GCSGS increased 48% and 59% compared with seedlings without inoculation, respectively. The inoculation of CBSB1 and CBSB1-GCSGS could increase the Cd and Pb extraction amounts of seedlings significantly compared with those without inoculation (P<0.05), the seedlings inoculated with CBSB1-GCSGS showed the highest Cd and Pb extraction amounts. It was concluded that the gcsgs gene introduced into Enterobacter sp. CBSB1 upgraded the phytoremediation efficacy of B. juncea. So the engineered Enterobacter sp. CBSB1-GCSGS showed potentials in remediation sites contaminated with complex contaminants by inoculating into remediating plants.

Enhancement of phytoremediation of Cd- and Pb-contaminated soils by self-fusion of protoplasts from endophytic fungus Mucor sp. CBRF59

The aim of this study was to isolate protoplasts from endophytic fungi and to carry out self-fusion of protoplasts for their enhancement of metal resistance. Self-fusant CBRF59T3 with resistance to 25 mM Cd(II) was constructed by self-fusion of inactivated protoplasts from Mucor sp. CBRF59. The inoculation of CBRF59 and CBRF59T3 improved significantly the availability of Cd(II) and Pb(II) in the soil. Compared with CBRF59, CBRF59T3 inoculation increased the content of water-soluble Cd(II) by 24%. The dry weight of rape inoculated with CBRF59 and CBRF59T3 was both higher than that of the uninoculation rape. Inoculation of CBRF59T3 further increased the dry weight of rape by 62% than CBRF59 in the higher Cd(II)-+Pb(II)-contaminated soil. Compared with CBRF59, CBRF59T3 inoculation increased the concentration of Cd(II) in rape shoots by 35-189% in Cd(II)- and Cd(II)-+Pb(II)-contaminated soils. The inoculation of CBRF59T3 also enhanced the translocation of Cd(II) from roots to shoots and increased the amount of extracted Cd(II) by rape. The results indicated that the mutants constructed by protoplast fusion is a feasible and efficient method to improve stress tolerance of uncharacterized fungi for phytoremediation of soils contaminated by heavy metals.

Effects of Cd, Pb, Zn, Cu-Resistant Endophytic Enterobactersp. CBSB1 and RhodotorulaSp. CBSB79 on the Growth and Phytoextraction of Brassica Plants in Multimetal Contaminated Soils

To survey the effects of endophytic Enterobacter sp. CBSB1 and Rhodotorula sp. CBSB79 resistant to Cd2+, Pb2+, Zn2+, and Cu2+ on the growth and phytoextraction of Brassica, the endophytes were isolated by surface- sterilized methods and characterized. The CBSB1 significantly increased 44.2% of the dry weight of Brassica napus in the multimetal contaminated soil (P < 0.05) and showed no effect or declined the dry weight of B. alboglabra, B. campestris ssp. chinensis var. cummunis, B. campestris ssp. chinensis var. utilis cv. Youqing12, B. campestris ssp. chinensis var. utilis cv. Lvbao701 plants. The dry weights of B. napus, B. campestris ssp. chinensis var. utilis, and B. alboglabra showed a significant increase when the CBSB79 was inoculated (P < 0.05). In general, inoculation with bacteria and yeast did not greatly alter the metal concentration in plant tissues. Compared to Enterobacter sp. CBSB1, the yeast Rhodotorula sp CBSB79 showed higher potentials to improve extraction efficacy of Cd, Pb, Zn, and Cu by Brassica seedlings in the field.

Seasonal and spatial diversity of microbial communities in marine sediments of the South China Sea

This study was conducted to characterize the diversity of microbial communities in marine sediments of the South China Sea by means of 16S rRNA gene clone libraries. The results revealed that the sediment samples collected in summer harboured a more diverse microbial community than that collected in winter, Deltaproteobacteria dominated 16S rRNA gene clone libraries from both seasons, followed by Gammaproteobacteria, Acidobacteria, Nitrospirae, Planctomycetes, Firmicutes. Archaea phylotypes were also found. The majority of clone sequences shared greatest similarity to uncultured organisms, mainly from hydrothermal sediments and cold seep sediments. In addition, the sedimentary microbial communities in the coastal sea appears to be much more diverse than that of the open sea. A spatial pattern in the sediment samples was observed that the sediment samples collected from the coastal sea and the open sea clustered separately, a novel microbial community dominated the open sea. The data indicate that changes in environmental conditions are accompanied by significant variations in diversity of microbial communities at the South China Sea.

Endophytic bacterial and fungal microbiota in sprouts, roots and stems of rice (Oryza sativa L.)

The objective of this study was to elucidate the endophytic microbiota in rice sprouts, roots, and stems, and their transmission in the plant development. Prior to DNA extraction, roots and stems were treated with 36% formaldehyde and 0.1M NaOH solutions to remove epiphytic bacterial whole 16S rRNA genes. Bacterial and fungal taxa in the sprout, root, and stem samples were analyzed using Illumina-based sequencing of the V3-V4 hyper variable regions of bacterial 16S rRNA genes and the ITS2 regions of fungal rRNA genes, respectively. Results showed that more diverse bacterial OTUs were detected in roots than in stems, while more diverse fungal OTUs were detected in stems than in roots. Compared with the endophytic microbiota in sprouts, the bacterial OTUs increased in roots but decreased in stems, whereas the fungal OTUs in both stems and roots decreased. Sprout-borne bacterial genera Sphingomonas and Pseudomonus, and fungal genera Fusarium, Pestalotiopsis, and Penicillium were detected in stems and roots. The coexistence of these indigenous bacterial and fungal taxa in sprouts, roots, and stems indicated their transmission during the development from sprouts to mature plants. The results from this study should be useful to better understand the plant-microbe interactions and to select suitable microbial taxa for rice production.