Endophytic bacteria reside within plant hosts without causing disease symptoms. In this study, 853 endophytic strains were isolated from aerial tissues of four agronomic crop species and 27 prairie plant species. We determined several phenotypic properties and found approximately equal numbers of gram-negative and gram-positive isolates. In a greenhouse study, 28 of 86 prairie plant endophytes were found to colonize their original hosts at 42 days postinoculation at levels of 3.5 to 7.7 log 10 CFU/g (fresh weight). More comprehensive colonization studies were conducted with 373 corn and sorghum endophytes. In growth room studies, none of the isolates displayed pathogenicity, and 69 of the strains were recovered from corn or sorghum seedlings at levels of 8.3 log 10 CFU/plant or higher. Host range greenhouse studies demonstrated that 26 of 29 endophytes were recoverable from at least one host other than corn and sorghum at levels of up to 5.8 log 10 CFU/g (fresh weight). Long-range dent corn greenhouse studies and field trials with 17 wild-type strains and 14 antibioticresistant mutants demonstrated bacterial persistence at significant average colonization levels ranging between 3.4 and 6.1 log 10 CFU/g (fresh weight) up to 78 days postinoculation. Three prairie and three agronomic endophytes exhibiting the most promising levels of colonization and an ability to persist were identified as Cellulomonas, Clavibacter, Curtobacterium, and Microbacterium isolates by 16S rRNA gene sequence, fatty acid, and carbon source utilization analyses. This study defines for the first time the endophytic nature of Microbacterium testaceum. These microorganisms may be useful for biocontrol and other applications.

Both the common bacterial blight (CBB) pathogen (Xanthomonas campestris pv. phaseoli) and X. fuscans subsp. fuscans, agent of fuscous blight, cause indistinguishable symptoms in common bean, Phaseolus vulgaris. Yield losses can exceed 40%. Lack of information about the specificity between X. campestris pv. phaseoli strains and major quantitative trait loci (QTL) or alleles conferring resistance makes the task of identifying genetic changes in host–pathogen interactions and the grouping of bacterial strains difficult. This, in turn, affects the choice of pathogen isolates used for germplasm screening and complicates breeding for CBB resistance. Common bean host genotypes carrying various sources and levels of resistance to CBB were screened with 69 X. campestris pv. phaseoli and 15 X. fuscans subsp. fuscans strains from around the world. Differential pathogenicity of the CBB pathogen was identified on the 12 selected bean genotypes. The X. fuscans subsp. fuscans strains showed greater pathogenicity than X. campestris pv. phaseoli strains having the same origin. African strains were most pathogenic. The largest variation in pathogenicity came from X. campestris pv. phaseoli strains that originated in Caribbean and South American countries. Pathogenic variation was greater within X. campestris pv. phaseoli than within X. fuscans subsp. fuscans strains. Implications for breeding for CBB resistance are discussed.

Clavibacter michiganensis subsp. nebraskensis (CMN) is a gram-positive bacterium and an incitant of Goss's bacterial wilt and leaf blight or "leaf freckles" in corn. A population structure of a wide temporal and geographic collection of CMN strains (n = 131), originating between 1969 and 2009, was determined using amplified fragment length polymorphism (AFLP) analysis and repetitive DNA sequence-based BOX-PCR. Analysis of the composite data set of AFLP and BOX-PCR fingerprints revealed two groups with a 60% cutoff similarity: a major group A (n = 118 strains) and a minor group B (n = 13 strains). The clustering in both groups was not correlated with strain pathogenicity. Group A contained two clusters, A1 (n = 78) and A2 (n = 40), with a linkage of 75%. Group A strains did not show any correlation with historical, geographical, morphological, or physiological properties of the strains. Group B was very heterogeneous and eight out of nine clusters were represented by a single strain. The mean similarity between clusters in group B varied from 13% to 63%. All strains in group B were isolated after 1999. The percentage of group B strains among all strains isolated after 1999 (n = 69) was 18.8%. Implications of the findings are discussed.

Bacterial wilt caused by Curtobacterium flaccumfaciens pv. flaccumfaciens was one of the more problematic diseases of dry bean (Phaseolus vulgaris L.) throughout the irrigated High Plains (Colorado, Nebraska, and Wyoming) in the 1960s and early 1970s, but has not been observed since that time. However, in August of 2003, plants exhibiting wilting and irregular, interveinal necrotic foliar lesions surrounded by a bright yellow border were found in three dry bean fields (market class Great Northern) in Scotts Bluff County, Nebraska. During 2004, plants exhibiting identical symptoms were additionally found occurring in more than 40 dry bean fields in western Nebraska. Affected fields were planted with dry bean from multiple market classes and seed sources, including yellow bean, Great Northern bean, and pinto bean, and incidence varied from trace levels to 80 to 90%. Isolations were made from leaf and stem tissues and seeds collected after harvest from infected plants, and all yielded slow-growing, creamy yellow or orange, fluidal colonies on nutrient broth-yeast extract medium. The bacterium was identified as C. flaccumfaciens pv. flaccumfaciens based on cell morphology (coryneformshaped motile rods), positive Gram stain and KOH reactions, fatty acid profiles, and BIOLOG (Hayward, CA) identifications. Great Northern (cv. Orion) plants were inoculated by bacterial suspensions (5 × 107 CFU/ml) injected into leaf axils adjacent to the first fully expanded trifoliolate and were incubated in the greenhouse under ambient conditions fluctuating between 24 and 35°C. Wilting symptoms developed 7 days after inoculation with foliar necrosis and yellowing symptoms appearing after 24 days. Identical bacterial colonies were reisolated from inoculated tissues, completing Koch's postulates. Although recent reports of wilt have been made in North Dakota (2) and western Canada (1) in 1995 and 2002, respectively, they were based only on the presence of discolored seeds observed in dockage from processing plants after harvest. To our knowledge, this report represents the first widespread observations of bacterial wilt from field infections in Nebraska in more than 30 years. References: (1) J. R. Venette et al. Plant Dis. 79:966, 1995. (2) T. F. Hsieh et al. Plant Dis: 86:1275, 2002.