Comparison of media for recovery of Verticillium dahliae from soil

Polygalacturonic acid (PGA) is an important constituent of Sorensen's NP-10 medium (NP-10) for estimating the population density of Verticillium dahliae in soil. Different types of PGA are available, but not all of them favor the growth of V dahliae. Unavailability of PGA sodium salt from orange (P-1879) has created an unprecedented problem for the quantification of microsclerotia (MS) of V dahliae in soil. The PGA from orange (P-3889) that is now available does not support the growth of V dahliae. Therefore, experiments were conducted to optimize the use of NP-10 prepared with P-3889 and various concentrations of NaOH. NP-10 with P-3889 amended with eight concentrations of NaOH were compared with NP-10 prepared from PGA sodium salt from orange (P-1879, now discontinued) and citrus (P-3850) along with cellophane and Na-pectate media for recovery of MS from soil and growth of V dahliae on the media. Seven soils were assayed for MS, and eight isolates of V dahliae were evaluated for growth and production of MS. Concentrations of NaOH >0.035N and <0.02N in NP-10 with P-3889 reduced mycelial growth, microsclerotial production, and recovery of MS from soils. Similarly, NP-10 with P-3850 alone, cellophane, and Na-pectate media had significantly reduced growth on media and recovery of V dahliae from soils. The NP-10 with P-3889 and 0.025N NaOH consistently yielded numbers of V dahliae MS from soil samples and supported the growth and production of MS similar to the NP-10 with P-1879. The medium developed in this study can serve as a direct replacement for the original NP-10 that was developed nearly three decades ago, an important component of which is no longer available.