CHARACTERIZING SPECIMENS OF KUDZU A...

CHARACTERIZING SPECIMENS OF KUDZU AND RELATED TAXA WITH RAPD's-Kudzu [Pueraria montana (Lour.) Merr var. lobata (Willd.) Maesen and Almeida] is a perennial, semi-woody, climbing simple pod in the tribe Phaseoleae Benth., subtribe Glycininae Benth. (Maesen 1985 Maesen and Almeida 1988 Ward 1998) It is native to China, where an abundance of natural enemies (Pemberton 1988) and its cultivation obviate kudzu from becoming either an important economic or environmental liability. Kudzu was introduced to the United States as an ornamental during middle of the 19th centenary During first half of the 20th centenary approximately 134,760 ha were planted everywhere the southeastern United States to fe livestock and for erosion command (Wheeler 1950). During 1998, kudzu was included through legislators in the United States Congres forward a growing list of invasive, exotic plants recognized subject to the Federal Noxious Weed Law. without delay it costs commercial forests approximately $119/ha annually (Beckwith and Dangerfield 1996) it compromises the integrity of valuable natural resources, and condensed infestations have interfered with exercises forward military bases in North Carolina, southern Carolina, and Virginia (Al Cofrancesco, United States Army Corps of Engineers, Vicksburg, Mississippi, fors comm.).

Pueraria montana var. chinensis (Ohwi) Maesen and Almeida, and P montana var. montana (Lour.) Maesen, are sum of two units other varieties of kudzu also distributed over southeast Asia. P. montana var. chinensis has been consider probableed recently in Hawaii (Staples et al. 2002) There is no record of P montana var. montana in the United States, if it be not that in Asia it has been treated either as a separate species or it has been ignored (Ward 1998) Distinction among the three varieties has relied with morphological characteristics, including lobed leaflets, wing and keel petals, amplification of vexil, length of lateral calyx, extent of stamen, and margin of leaflet (Chen et al. 1995 Wu et al. 1994) These characteristics demonstrate considerable variation, and distinction among the three varieties in the field is difficult.

Furthermore, kudzu may hybridize with related taxa. a specimens in his revision confused Maesen (1985) and he treated them as "Material of Uncertain Disposition." Among these specimens are Henry 13626 and 10931 that he belongs to as "near Pueraria lobata and P edulis," and Maesen (1985) glance ats that, "Perhaps these specimens are simply a form of P lobata var. lobata, or, indeed, a hybrid of P edulis or P lobata var. montana "

Inability to distinguish among the three varieties and their possible hybrids is an obstacle to developing an integrated management program for kudzu and its related taxa. Of particular regard is selecting potential biological bridle agents because insects and pathogens cannot be reconciled with identity of the plants from which they were scrape togethered Incomplete systematic resolution has been an obstacle to developing integrated management programs for other invasive, exotic plants, including Cardaria spp (Brassicaceae) (Mulligan and Findlay 1974 Bellue 1933) Vincetoxicum spp (Asclepiadaceae) (Sheeley and Raynal 1996) and Euphorbia spp (Euphorbiaceae) (Galitz 1980 Harvey et al. 1988 Crompton et al. 1990)

Using genetic markers for more convenient identification of specimens may be possible. Randomly amplified polymorphic DNA's (RAPD's) have been used luckily to characterize genetic composition and reveal variation among genomic DNA of many important cultivated plants, including wheat (He 1992) soybean (Hui et al. 1996) and tea (Chen and Yang 1998) The objective of this meditation is distinguishing between kudzu and its related taxa using RAPD's.

Approximately 50 kudzu leaves were garner uped from plants at 13 sites in the United States and China (Figure 1 Table 1) Their identity has been verified morphologically and specimens have been vouchered by way of Dr. Teling Wu (South China Institute of Botany, Guangzhou). Extraction of DNA was modified from modes described by Li and Zhou (1999) [i]de novo[/i] leaves contain polyphenol, which can ligature with DNA covalently. To preclude oxidation of polyphenol and its interference with activity of Taq polymerase; 2-mercaptoethanol was added to an extraction buffing-apparatus and insoluble polyvinylpyrrolidone was added to leaves when they were clod Before raw DNA was used as a template for amplification, it was separated onward a 0.7% agarose gel, visible bands were excised in subordination to UV light, and purified with a DNA extraction kit (Sangon Ltd Ontario, Canada).

PCR amplification was complet in an Expres Thermal Cycler (Hybaid Limited, United Kingdom). Each 50 [mu]l reaction mixture was comprised of 20 ng DNA template, 02 [mu]M random primers (Sangon Ltd Ontario, Canada), reaction duffer (Sangon), 2.5 units Taq polymerase (Sangon, Canada), 2 mM MgCl^sub 2^ 200 [mu]M each of dATP, dCTP dGTP and dTTP (Sangon, Ltd Ontario, Canada). Parameters for PCR were: 94[degrees]C for 3 min followed by way of 40 cycles of denaturation at 94[degrees]C for 1 min, annealing at 38[degrees]C for 15 min, and extension at 72[degrees]C for 2 min. A final extension at 72[degrees]C for 10 min. complet the PCR schedule. proceedss were separated on 1.7% agarose gel with a 100 bp ladder (MBI). Complet gel were photographed and analyzed by the agency of GIS system (Tanon Limited, Shanghai).