Phytophthora species were surveyed by collecting soil samples and placing bait leaves in selected streams during June-October in the years 2005, 2006 and 2010 at three sites in oak forests in Diqing Tibetan Autonomous Prefecture of NW Yunnan province, China. Seventy-three isolates of Phytophthora spp. were recovered from 135 baited leaf samples and 81 soil samples. Eight Phytophthora species were identified by observation of morphological features and ITS1-5.8S-ITS2 rDNA sequence analysis. The eight taxa included two well-known species P. gonapodyides and P. cryptogea, two recently described species P. gregata and P. plurivora, two named but as yet undescribed taxa, P. taxon PgChlamydo and P. taxon Salixsoil, and two previously unrecognized species, Phytophthora sp.1 and P. sp.2. The most numerous species, P. taxon PgChlamydo, and the second most abundant species, P. taxon Salixsoil, were recovered at all three sites. Phytophthora cryptogea was detected only once at site Nixi. Phytophthora gregata and P. sp.2 were isolated from a stream only at site Bitahai, while the other three species were each found at two sites. Phylogenetic analysis revealed that the isolates belonged to three ITS clades, one species including six isolates in clade 2, six species including 66 isolates in clade 6 and one species in clade 8. There was a relatively rich species and genetic diversity of Phytophthora detected in the investigated regions where the forest biotic and abiotic factors affecting the growth and evolution of Phytophthora populations were diverse.

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Acidovorax citrulli causes seedling blight and bacterial fruit blotch (BFB) of cucurbits, a serious disease threat to cucurbit seed and fruit production worldwide. Despite its economic importance, current knowledge on the biology and pathology of A. citrulli is limited. This work studies a mini-Tn5 mutant, MJ22-3, that showed reduced virulence on melon seedlings. The gene disrupted by Tn5 was identified as leuB, which encodes a putative 3-isopropylmalate dehydrogenase, an enzyme that appears to be essential for leucine biosynthesis. The leuB gene in-frame deletion mutant MDleuB, as well as the Tn5-inserted mutant MJ22-3, failed to grow in minimal medium without leucine and exhibited reduced swimming motility; however, the mutations had no effect on biofilm formation or induction of the hypersensitive response on tobacco (Nicotiana tabacum). When inoculated at a low cell density (10 4 CFU mL À1), the leuB gene mutants showed a significantly slower growth rate and attenuated virulence on melon cotyledons compared to the wildtype MH21. However, inoculations with the mutants at high concentration (10 8 CFU mL À1 ), or low concentration (10 4 CFU mL À1 ) inoculations supplemented with 0Á01% leucine resulted in growth rates and disease severities similar to that of the wildtype MH21. The results suggest that leucine biosynthesis is essential for both in vivo growth and full virulence of A. citrulli MH21 on melon seedlings.

Phytoplasmas were detected in Sophora japonica cv. golden and Robinia pseudoacacia with diseased branches of witches'-broom collected in Haidian district, Beijing, China. Phytoplasma cells were observed in phloem sieve elements of symptomatic S. japonica cv. golden by transmission electron microscopy. The presence of phytoplasmas was further confirmed by sequence determination of partial gene sequences of 16S rDNA, rp (ribosomal protein) and secY. Phylogenetic trees and virtual restriction fragment length polymorphism (RFLP) analyses indicated that the phytoplasmas causing S. japonica cv. golden witches'-broom (SJGWB) and R. pseudoacacia witches'-broom (RPWB) belong to the 16SrV (elm yellows) group, and they are most closely related to subgroup 16SrV-B, rpV-C and secYV-C jujube witches'-broom (JWB) phytoplasma. Comparative analyses indicated that the phytoplasma of RPWB was closer to the JWB and that R. pseudoacacia might serve as an alternative host plant of JWB phytoplasma.

Bumald spirea (Spiarea bumalda Burv.) is an important ornamental tree widely grown in northern China. In August of 2006, spirea plants exhibiting symptoms of witches'-broom, stunting, yellowing, and shoot dieback were found at an incidence of 5 to 15% in Qingzhou City, Shandong Province, China. Total DNA was extracted separately from 0.1 g of phloem tissue from leaf midribs and stems of six symptomatic and six asymptomatic plants with a modified cetyltriethylammonium bromide (CTAB) method (3). Resulting DNA samples were analyzed for phytoplasma DNA by a nested PCR assay using phytoplasma universal 16S rDNA gene primer pairs R16mF2/R16mR1 and R16F2n/R16R2 (2). These primers amplified 1.5- and 1.2-kb products, respectively, from DNA of all symptomatic plants only. Restriction fragment length polymorphism (RFLP) analysis of the 1.2-kb 16S rDNA product using enzymes AluI, MseI, and HhaI indicated that all symptomatic plants contained a group 16SrI (aster yellows group) subgroup B (16SrI-B) phytoplasma strain (4). A 16S rDNA sequence derived from this strain (GenBank Accession No. EF176608) was most similar (99.8 and 99.6%) to those of severe aster yellows (GenBank Accession No. M86340) and Maryland aster yellows (GenBank Accession No. AF322644) phytoplasmas, respectively, thereby confirming strain identity based on RFLP analysis. A phytoplasma (Spiarea stunt phytoplasma, GenBank Accession No. AF190228), which belongs to X-disease group (16SrIII), was reported to infect spirea and probably be lethal to S. tomentosa in New York (1,4). The phytoplasma reported here shared low identity (90.8%) with Spiarea stunt phytoplasma, but also caused dieback of spirea shoots. The epidemiology and economic impact of this disease need further intensive investigation. To our knowledge, this is the first report of spirea witches'-broom disease and of its association with a subgroup 16SrI-B phytoplasma in China. References: (1) H. M. Griffiths et al. Can. J. Plant Pathol. 16:255, 1994. (2) D. E. Gundersen and I.-M. Lee. Phytopathol. Mediterr. 35:144, 1996. (3) Y. Qi et al. Biotechnol. Bull. 4:44, 2004. (4) The IRPCM Phytoplasma/Spiroplasma Working Team-Phytoplasma Taxonomy Group. Int. J. Syst. Bacteriol. 54:1243, 2004.

The green mirid bug Apolygus lucorum is a regional pest of multiple crops in northern China, and the survival and development of diapausing eggs during winter plays an important role in the population dynamics of this species. The effect of water on the survival and development of A. lucorum eggs was investigated using laboratory-induced diapause. Diapausing eggs were exposed to various humidity regimes under three conditions: (1) termination of diapause with exposure to warm long-day (WLD) conditions (i.e., 26 ± 1°C and 75 ± 5% relative humidity (RH) under a photoperiod of 16 hours light and 8 hours dark), (2) termination of diapause by chilling at 4°C, or (3) during the post-diapause stage, i.e., from transfer to WLD conditions after chilling, until the hatching of nymphs. The results indicate that water availability is crucial for the post-diapause resumption of development of A. lucorum. However, exposure to excessive moisture was detrimental, as indicated by a decrease in diapause termination rate and a prolonged pre-hatching period of diapausing eggs, compared to limited moisture conditions. This implies that both too dry and too humid environmental conditions would suppress survival and postpone hatching of overwintered A. lucorum eggs, and might explain why this pest has not caused severe damage in either southern or western China where the respective climates are very humid or dry.

Bacterial contamination and the spread of antibiotic-resistant bacteria demand alternate methods to deal with bacterial infections. With particular advantages, photodynamic therapy (PDT) is a promising approach. As a kind of photosensitizer for PDT, light-induced antibacterial compounds like oligo-p-phenylene-ethynylenes (OPEs) have been widely investigated while these studies mainly focus on OPEs with quaternary ammonium salts. In our previous study, OPEs with tertiary amino groups (T-OPEs) were reported to exhibit a better antibacterial activity than the corresponding quaternary ammonium salts, which make it important to develop T-OPEs and further investigate their structure–activity relationship. Additionally, the terminal structure of the reported OPEs mainly consists of quaternary ammonium salts or tertiary amino groups, which could not be linked to other materials. Thus, to develop more effective and multifunctional antibacterial agents, we designed and synthesized four unsymmetrical OPEs having terminal amino groups, which could be linked to other functional units by covalent bonds. Their antibacterial activity against Gram-positive and Gram-negative bacteria and the mechanism have been investigated. The OPEs showed effective biocidal activity under fiber light irradiation, and no dark killing was observed. The mechanism study indicates that OPEs could penetrate and perturb the cell membrane and generate ROS under light irradiation, both of which could influence their antibacterial activity. The penetrating ability of OPEs is partly dependent on their lipophilicity and the structure and composition of the cell membrane.

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