Isolation and transformation ofTrichoderma virideprotoplasts

2004 ◽  
Vol 1 (2) ◽  
pp. 67-72 ◽  
Author(s):  
Liu Shi-Wang ◽  
Wang Zheng-Yi ◽  
Guo Ze-Jian
Keyword(s):  
Resistance Gene ◽  
Phosphate Buffer ◽  
Culture Medium ◽  
Trichoderma Viride ◽  
Protoplast Isolation ◽  
Hygromycin Resistance ◽  
Protoplast Yield ◽  
Hygromycin Resistance Gene

AbstractThe conditions for protoplast isolation and regeneration fromTrichoderma viridewere studied. Protoplasts were optimally isolated when mycelia ofT. viridethat had been cultured for 24 h were digested with 4 mg/ml Glucanex in phosphate buffer (pH 6.98) for 4 h at 30°C, resulting in a protoplast yield of 4.7×107cfu/mg. The maximum regeneration ratio (14.5%) was obtained in mycelia culture medium containing 0.3 mol/l KCl and 0.3 mol/l inositol. In addition, a plasmid pCSSNCC1 carrying a hygromycin resistance gene and an elicitor-producing gene was transformed intoT. virideprotoplasts, with an efficiency of 1–2 transformants/μg DNA. The hygromycin-resistant transformants were determined by PCR and the elicitor protein was detected by ELISA. The results indicate that the elicitor protein was expressed stably inT. viride.

The Transformation in Trichoderma viride Protoplasts by Restriction Enzyme Mediated Integration (REMI)

2012 ◽  
Vol 518-523 ◽  
pp. 545-548
Author(s):  
Yu Ru You ◽  
Shi Wang Liu ◽  
Yuan Feng Wu ◽  
Jun Huang ◽  
Gong Nian Xiao ◽  
...  
Keyword(s):  
Resistance Gene ◽  
Restriction Enzyme ◽  
Phosphate Buffer ◽  
Trichoderma Viride ◽  
Hygromycin B ◽  
Hygromycin B Resistance ◽  
Protoplast Yield ◽  
Preliminary Identification ◽  
Exogenous Gene

The transformation conditions of the protoplasts from Trichoderma viride mediated by restriction enzyme were studied in this paper. The optimum generation conditions of protoplasts were as followed: 8 mg/ml glucanex was added into the phosphate buffer (pH 6.98), the mycelial that cultured for 24 hr was hydrolyzed for 4 hr at 30°C under 40 r/min shaking speed. The protoplast yield was 4.7×107 cfu/mg. The regeneneration rate of protoplast was 14.5% on CM medium contained 0.3 mol/L KCl and 0.3 mol/L Inositol. Transformants were obtained by transfering hygromycin B resistance gene into T. viride by restriction enzyme mediated integration (REMI), The preliminary identification of the transformants indicated that the exogenous gene had been integrated into T. viride genome.

A chimaeric hygromycin resistance gene as a selectable marker in plant cells

1985 ◽  
Vol 5 (5) ◽  
pp. 299-302 ◽  
Author(s):  
Peter J. M. van den Elzen ◽  
Jeffrey Townsend ◽  
Kathleen Y. Lee ◽  
John R. Bedbrook
Keyword(s):  
Resistance Gene ◽  
Selectable Marker ◽  
Plant Cells ◽  
Hygromycin Resistance ◽  
Hygromycin Resistance Gene

A mutated hygromycin resistance gene is functional in the n -alkane-assimilating yeast Candida tropicalis

2000 ◽  
Vol 173 (3) ◽  
pp. 187-192 ◽  
Author(s):  
Akihiro Hara ◽  
Mitsuyoshi Ueda ◽  
Satoru Misawa ◽  
Toru Matsui ◽  
Keizo Furuhashi ◽  
...  
Keyword(s):  
Resistance Gene ◽  
Candida Tropicalis ◽  
Hygromycin Resistance ◽  
Hygromycin Resistance Gene

scholarly journals Hygromycin Resistance Gene Cassettes for Vector Construction and Selection of Transformed Rice Protoplasts

1991 ◽  
Vol 97 (2) ◽  
pp. 832-835 ◽  
Author(s):  
Zhenwei Zheng ◽  
Akio Hayashimoto ◽  
Zhijian Li ◽  
Norimoto Murai
Keyword(s):  
Resistance Gene ◽  
Hygromycin Resistance ◽  
Gene Cassettes ◽  
Vector Construction ◽  
Hygromycin Resistance Gene ◽  
Selection Of

Transformation of Aspergillus nidulans with the hygromycin-resistance gene, hph

Gene ◽  
1987 ◽  
Vol 57 (1) ◽  
pp. 21-26 ◽  
Author(s):  
D. Cullen ◽  
S.A. Leong ◽  
L.J. Wilson ◽  
D.J. Henner
Keyword(s):  
Aspergillus Nidulans ◽  
Resistance Gene ◽  
Hygromycin Resistance ◽  
Hygromycin Resistance Gene

scholarly journals Stability and modulated expression of a hygromycin resistance gene integrated in Botrytis cinerea transformants

2006 ◽  
Vol 154 (2) ◽  
pp. 187-193 ◽  
Author(s):  
Walid Hamada ◽  
Marie-Christine Soulié ◽  
Pierrette Malfatti ◽  
Gilbert Bompeix ◽  
Martine Boccara
Keyword(s):  
Botrytis Cinerea ◽  
Resistance Gene ◽  
Hygromycin Resistance ◽  
Hygromycin Resistance Gene

scholarly journals Transfer of a Supernumerary Chromosome Between Vegetatively Incompatible Biotypes of the Fungus Colletotrichum gloeosporioides

Genetics ◽  
1998 ◽  
Vol 150 (4) ◽  
pp. 1459-1466 ◽  
Author(s):  
Chaozu He ◽  
Anca G Rusu ◽  
Agnieszka M Poplawski ◽  
John A G Irwin ◽  
John M Manners
Keyword(s):  
Resistance Gene ◽  
Colletotrichum Gloeosporioides ◽  
Mixed Cultures ◽  
Marker Genes ◽  
Hygromycin Resistance ◽  
Antibiotic Resistant ◽  
Selectable Marker Genes ◽  
Tropical Legumes ◽  
Hygromycin Resistance Gene ◽  
Biotype B

Abstract Two biotypes (A and B) of Colletotrichum gloeosporioides infect the tropical legumes Stylosanthes spp. in Australia. These biotypes are asexual and vegetatively incompatible. However, field isolates of biotype B carrying a supernumerary 2-Mb chromosome, thought to originate from biotype A, have been reported previously. We tested the hypothesis that the 2-Mb chromosome could be transferred from biotype A to biotype B under laboratory conditions. Selectable marker genes conferring resistance to hygromycin and phleomycin were introduced into isolates of biotypes A and B, respectively. A transformant of biotype A, with the hygromycin resistance gene integrated on the 2-Mb chromosome, was cocultivated with phleomycin-resistant transformants of biotype B. Double antibiotic-resistant colonies were obtained from conidia of these mixed cultures at a frequency of approximately 10-7. Molecular analysis using RFLPs, RAPDs, and electrophoretic karyotypes showed that these colonies contained the 2-Mb chromosome in a biotype B genetic background. In contrast, no double antibiotic colonies developed from conidia obtained from mixed cultures of phleomycin-resistant transformants of biotype B with biotype A transformants carrying the hygromycin resistance gene integrated in chromosomes >2 Mb in size. The results demonstrated that the 2-Mb chromosome was selectively transferred from biotype A to biotype B. The horizontal transfer of specific chromosomes across vegetative incompatibility barriers may explain the origin of supernumerary chromosomes in fungi.

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