[Preliminary report on the use of total lumpectomyconical remnant gastric - esophagus side overlap anastomosis in radical resection of Siewert type II proximal gastric cancer].

Objective: There is no standard method for esophageal remnant gastric reconstruction for proximal gastrectomy. Reflux esophagitis caused by esophagogastrostomy remains a difficult surgical problem. To report the preliminary surgical results of novel esophagus-conical remnant gastric side overlap anastomosis (CGEO) , with particular emphasis on postoperative esophageal reflux. Methods: In June 2022, we developed a novel CGEO for laparoscopic proximal gastrectomy on two patients with Siewert type II esophagogastric junction adenocarcinoma. Surgical procedures for CGEO: (1) Laparoscopic proximal gastrectomy and preparation of conically shaped gastric remnant; (2) Determining anastomotic site of residual stomach and esophagus; (3) Side-to-side anastomosis of right esophageal wall to anterior of conical gastric remnant; (4) Valvuloplasty of esophageal stump. Results: Case 1 was a 71-year-old man with an operation time of 305 minutes and was successfully discharged from the hospital on the 9th day after surgery, and the postoperative pathology was T3N0M0. Case 2 was an 82-year-old man with an operation time of 325 minutes. He was discharged on the 10th day after surgery. In both cases, only mild esophageal mucosal changes were seen in gastroscopy, there were no obvious symptoms of esophageal reflux. There was also no significant weight change at half a year after operation. Conclusion: CGEO is moderately safe in radical surgery for proximal gastric cancer, and may have a preventive effect on the occurrence of postoperative esophageal reflux, but long-term results need to be confirmed by further studies with follow-up.

First Report of a Pestalotiopsis sp. Causing Leaf Spot of Blueberry in China.

In August 2006, leaf spots were observed on half-high blueberry (Vaccinium corymbosum) in a plant nursery in Dalian, China. The symptomatic potted 1-year-old blueberry plants were located in parts of a plant nursery with poor ventilation. The primary symptom was a leaf spot, 0.4 to 0.8 cm in diameter, with brown margins that enlarged and coalesced. Mycelium grew from symptomatic and green leaf tissue removed from the margin of a necrotic leaf spot. Plant tissues were surface disinfested with 0.1% mercuric chloride for 3 min and 70% ethyl alcohol for 30 s before plating onto potato dextrose agar. The resulting colonies were white with a regular margin and a rough surface. The cultures were covered with black and globular acervuli with a diameter of 100 to 200 µm. The base of each conidiophore was swollen and globose with phialides growing from the apical end. Mature conidia were straight to fusiform, measuring 19.0 to 27.5 × 6.3 to 9.2 µm, and five-celled with the three middle cells brown and darker than the end cells. The apical cell was triangular and hyaline with three simple setulae that were 17.2 to 29.7 µm long. The base cell terminated in a point 4.0 to 8.6 µm long. Koch's postulates were fulfilled for the fungus by spray inoculating two healthy young plants with 2 × 105 conidia per ml of sterile distilled water. As a control, two similar plants were sprayed with sterile water. Plants were placed inside plastic bags to maintain humidity and incubated in a growth chamber at 26°C under fluorescent light for 14 h and at 20°C in darkness for 10 h. After 3 days, the plastic bags were removed and plants were maintained under the same conditions. More than 20 days after inoculation, symptoms on inoculated plants were similar to those previously described in the nursery. Control plants did not show any symptoms. Cultures isolated from the lesions were similar to those isolated previously from plants in the nursery. The morphological descriptions and measurements were similar to Pestalotiopsis clavispora (1). The 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA and partial ß-tubulin gene were amplified from DNA extracted from single-spore cultures using the ITS1/ITS4 and T1/Bt2b primers (2) respectively, and sequenced (GenBank Accession Nos. EF119336 and EF152585). The ITS sequences were most similar to the ITS regions of P. clavispora TA-8 (98%; GenBank Accession No. AY924264), P. clavispora TA-6 (98%; GenBank Accession No. AY924263), and P. clavispora PSHI 2002 Endo 389 (96%; GenBank Accession No. AY682929). The partial ß-tubulin gene sequence was identical to Pestalotiopsis sp. isolate PSHI 2004 Endo 86 (100%; GenBank Accession No. DQ657901). The morphology and sequence data support the identity of the causal fungus as P. clavispora. To our knowledge, this is the first report on the presence of a Pestalotiopsis sp. causing a disease of blueberry in China. References: (1) E. F. Guba. Monograph of Monochaetia and Pestalotia. Harvard University Press, Cambridge, MA, 1961. (2) W. Tao et al. Mol. Cell Biol. 27:689, 2007.

First Report of Alternaria tenuissima Causing Disease on Blueberry in China.

During September 2006, disease symptoms were observed on mature highbush blueberry (Vaccinium corymbosum L.) cvs. Bluecrop and Covoille in a blueberry commercial field in Dalian, China. The maximum and minimum rainfalls in June to September are 3,111.9 and 1,745.6 ml, respectively. The highest temperature during the summer is 35.3°C and relative humidity may achieve 90%. Circular to irregular, light brown-to-gray leaf spots with brownish red borders, initially 3 to 7 mm in diameter, enlarged and coalesced. Reddish, circular spots appeared on stems, developing small, insignificant cankers. A fungus was recovered on potato dextrose agar (PDA, pH nature) from the margin of necrotic leaf spots. Morphological traits of the strain that developed from a single-spore culture were as follows: colonies were regular and flat, with a rough upper surface that peripherally was olive-green with a black center and dull white spots; short conidiophores arising singly and measuring 81.6 to 163.2 × 4.1 to 8.2 µm; conidia was abundant, ovoid, and obclavate muriformly septate, which horizontal and vertical septations varied from 1 to 6 and 0 to 2, respectively, and its size varied from 26 to 48.8 × 9.7 to 16.3 µm with an average beak length of 9.6 µm, and sporulation pattern is budding. Conidia derived from conidiophores. Koch's postulates were fulfilled for the isolates by spray inoculating two healthy mature plants with 2 × 105 conidia per ml homogenized in sterile water. As a control, two plants were sprayed with sterile water. Plants were placed inside plastic bags to maintain humidity and incubated in a growth chamber at 26°C under fluorescent light for 14 h and 20°C in darkness for 10 h. After 2 days, the plastic bags were removed and plants were maintained under the same conditions for 30 days. Symptoms on inoculated plants were similar to those previously observed. Symptoms were not observed on control plants. Cultures isolated from inoculated plants had the same morphological traits as those that were isolated previously from the field plants. The morphological descriptions and measurements were similar to Alternaria tenuissima (2). The 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA and partial cds histone gene were amplified from DNA extracted from single-spore cultures using the ITS1/ITS4 and H3-1a/H3-1b primers, respectively, and sequenced (GenBank Accession No. EF031053) (1,3). The ITS sequence was identical to the ITS regions of A. tenuissima strain EGS34-015 (100%; GenBank Accession No. AY751455), the partial cds histone gene sequence was similar to A. tenuissima isolate MA6 (99%; GenBank Accession No. AF404634). The morphology, secondary conidiation, and sequences of ITS and partial cds histone gene identify the causal fungus as A. tenuissima. To our knowledge, this is the first report on the presence of A. tenuissima affecting blueberry plants in China. References: (1) J. C. Kang et al. Mycol. Res. 106:1151, 2002. (2) E. G. Simmons. Mycotaxon 70:325, 1999. (3) T. J. White et al. Pages 315-322 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

First Report of Blueberry Leaf Spot Caused by Cylindrocladium colhounii in China.

During late July and early August of 2005, leaf spot symptoms were observed in a blueberry nursery at a plantation in Dalian, which to our knowledge, lies within the largest blueberry-production area in China. Symptoms were observed primarily on lowbush species, for example Blomidon, as well as half-highbush cultivars. A slow-growing, white mycelium from the margin of necrotic leaf spots was recovered on potato dextrose agar (PDA). The following morphological traits were observed: erect conidiophores that branch twice and were terminated in a stiped, clavate phialide; hyaline, cylindrical, four-celled conidia; and globose, reddish brown, aggregated chlamydospores. Conidiophores (including stipes and terminal phialides) were 305 to 420 × 5 to 9 µm; primary branches were 9 to 45 × 5 to 6.3 µm; secondary branches were 9 to 17.3 × 3.1 to 4.5 µm; phialides were 7.8 to 17.5 × 2.5 to 6 µm; stipes (from the highest branch area to vesicle) were 150 to 270 µm long; and vesicles were 13 to 30 × 2 to 4.5 µm. Conidia were 50 to 72 × 4 to 5.5 µm. Chlamydospores were 15 to 20 µm in diameter. Koch's postulates were fulfilled by spray inoculating two healthy cultivars with conidiophores homogenized in axenic water. As a control, two healthy plants were sprayed with axenic water. Plants were placed inside plastic bags to maintain humidity and incubated in a growth chamber at 26°C under fluorescent light for 14 h and 20°C in darkness for 10 h. After 2 days, the plastic bags were removed and plants were maintained under the same conditions. After 4 days, small-to-medium brown spots with purplish margins were observed on the adaxial side of leaves from inoculated plants, but not from control plants. Fungi isolated from these lesions had the same morphological traits as the ones isolated previously from field plants. The morphological descriptions and measurements were similar to Cylindorocladium colhounii (2). The 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA and the ß-tubulin gene were amplified from DNA extracted from single-spore cultures using the ITS1/ITS4 primers and T1/Bt2b primers, respectively, and sequenced (1). The ITS and ß-tubulin gene sequences were similar to C. colhounii STE-U 1237 (99%; GenBank Accession No. AF231953) and C. colhounii STE-U 705 (99%; GenBank Accession No. AF231954), respectively. The morphology, secondary conidiation, and sequences of ITS and ß-tubulin gene identify the causal fungus as C. colhounii. To our knowledge, this is the first report of C. colhounii on blueberry in China or in the world. References: (1) P. W. Crous et al. Can. J. Bot. 77:1813, 1999. (2) T. Watanabe. Page 222 in: Dictorial Atlas of Soil and Seed Fungi. CRC Press, Inc., Boca Raton, Fl, 1994.

First Report of Sweet potato leaf curl virus in China.

During the 2004 growing season in the Liaoning Province in China, where there was large population of whiteflies, several sweet potato (Ipomoea batatas) breeding lines showed leaf curl symptoms. A survey was conducted to determine the incidence of Sweet potato leaf curl virus (SPLCV) in China. Sixteen plants were collected and stem scions from those plants were graft inoculated to Ipomoea nil. Three weeks later, the indicator developed symptoms of leaf curling, interveinal chlorosis, and stunting. Total nucleic acid was extracted from young leaves of sweet potato and then evaluated using polymerase chain reaction (PCR). Primers, developed by Briddon and Markham (1) and used as universal primers for amplification of the geminivirus DNA fragment, were BM-V (5'-KSG GGT CGA CGT CAT CAA TGA CGT TRT AC-3') and BM-C (5'-AAR GAA TTC ATK GGG GCC CAR ARR GAC TGG C-3'). Amplified fragments with BM primers theoretically should have sizes almost equal to the full length of the DNA A component of the bipartite genome (2). Expected DNA fragments of 2.8 kb that contained the AV1, AV2, AC1, AC2, AC3, and AC4 open reading frames were obtained from symptomatic, but not from symptomless (uninfected) plants. The 2.8-kb fragments obtained by amplification were purified and cloned into the PMD18-T vector. Recombinant plasmids were then transformed into competent cells of Escherichia coli strain DH5(. The fragment was sequenced (GenBank Accession No. DQ512731), and nucleotide sequence of corresponding regions were compared with a published sequence of SPLCV available in GenBank (Accession No. AF104036). The AC4 and AC2 genes showed the highest (92%) and the lowest (83%) identity, respectively. This virus has been reported in the United States, Taiwan, Japan, and Peru. To our knowledge, this is the first report of the natural occurrence of SPLCV in China. References: (1) R. W. Briddon and P. G. Markham. Mol. Biotechnol. 1:202, 1994. (2) M. Onuki and K. Hanada. Ann. Phytopathol. Soc. Jpn. 64:116, 1998.