TY - JOUR
T1 - Identification and molecular characterisation of new Mexican isolates of Pochonia chlamydosporia for the management of Meloidogyne spp.
AU - Medina-Canales, Ma G.
AU - Rodríguez-Tovar, A. V.
AU - Manzanilla-López, R. H.
AU - Zúñiga, G.
AU - Tovar-Soto, A.
N1 - Funding Information:
We thank the Consejo Nacional de Ciencia y Tecnología (CONACYT) for the financial support provided through a fellowship for graduate studies. We also thank the financial support granted by the Secretaría de Investigación y Posgrado del IPN to carry out the present research as part of the projects SIP-IPN20100663 and SIP-IPN 20110658. Dr Leopoldo Hidalgo-Díaz of CENSA (Centro Nacional de Sanidad Agropecuaria, Cuba) for making available the Pc392 isolate for its use in the present study through the Rothamsted Research collection. Finally, we thank Dr. Nestor Pérez and the Biologist María Esther Sanchez Espíndola for valuable contributions to this work.
PY - 2014/1
Y1 - 2014/1
N2 - New Mexican isolates of the nematophagous fungus Pochonia chlamydosporia were obtained from nematode infested fields in the vegetable growing area of Tepeaca Valley, Puebla State, Mexico. Based on macro and microscopic morphology, seven 'putative' P. chlamydosporia isolates were selected and the DNA extracted for polymerase chain reaction (PCR). Three new isolates of P. chlamydosporia were identified: Pcp2, Pcp21 and Pcp31. The amplification reaction of the internal transcribed spacer (ITS) region revealed a 650 bp amplicon which was used in a maximum likelihood phylogenetic inference analysis. Three groups were recovered in the tree topology, supported by a > 90% bootstrap value. Nucleotide identity values were > 83.6% between the test sequences and the reference sequence. In addition, using specific primers for two existing varieties of P. chlamydosporia, restriction fragment length polymorphism on the ITS products in conjunction with the phylogenetic inferences and the molecular test for detection of P. chlamydosporia vcp1 gene, it was found that all three isolates belong to a new variety which we have named P. chlamydosporia var. mexicana. We compared the chlamydospore production rate, rhizosphere colonisation and egg parasitism percentages of the three native isolates in Meloidogyne spp. with a reference isolate (Pc10). Native isolates produced > 1×106 chlamydospores/50 g of substrate (of which more than 80% were viable), colonised > 80% of the rhizosphere, and parasitised > 60% of Meloidogyne incognita and Meloidogyne arenaria eggs. Meloidogyne hapla egg parasitism was < 60%. Isolates Pcp2 and Pcp21 were identified as potential biological control agents of Meloidogyne spp. to be tested further in greenhouse and field tests.
AB - New Mexican isolates of the nematophagous fungus Pochonia chlamydosporia were obtained from nematode infested fields in the vegetable growing area of Tepeaca Valley, Puebla State, Mexico. Based on macro and microscopic morphology, seven 'putative' P. chlamydosporia isolates were selected and the DNA extracted for polymerase chain reaction (PCR). Three new isolates of P. chlamydosporia were identified: Pcp2, Pcp21 and Pcp31. The amplification reaction of the internal transcribed spacer (ITS) region revealed a 650 bp amplicon which was used in a maximum likelihood phylogenetic inference analysis. Three groups were recovered in the tree topology, supported by a > 90% bootstrap value. Nucleotide identity values were > 83.6% between the test sequences and the reference sequence. In addition, using specific primers for two existing varieties of P. chlamydosporia, restriction fragment length polymorphism on the ITS products in conjunction with the phylogenetic inferences and the molecular test for detection of P. chlamydosporia vcp1 gene, it was found that all three isolates belong to a new variety which we have named P. chlamydosporia var. mexicana. We compared the chlamydospore production rate, rhizosphere colonisation and egg parasitism percentages of the three native isolates in Meloidogyne spp. with a reference isolate (Pc10). Native isolates produced > 1×106 chlamydospores/50 g of substrate (of which more than 80% were viable), colonised > 80% of the rhizosphere, and parasitised > 60% of Meloidogyne incognita and Meloidogyne arenaria eggs. Meloidogyne hapla egg parasitism was < 60%. Isolates Pcp2 and Pcp21 were identified as potential biological control agents of Meloidogyne spp. to be tested further in greenhouse and field tests.
KW - Pochonia chlamydosporia var. mexicana n. var.
KW - nematophagous fungi
KW - phylogenetic tree
KW - rhizoplane
KW - root-knot nematode
UR - http://www.scopus.com/inward/record.url?scp=84889027908&partnerID=8YFLogxK
U2 - 10.1080/09583157.2013.838623
DO - 10.1080/09583157.2013.838623
M3 - Artículo
SN - 0958-3157
VL - 24
SP - 1
EP - 21
JO - Biocontrol Science and Technology
JF - Biocontrol Science and Technology
IS - 1
ER -