Change in nuclear DNA content and pollen size with polyploidisation in the sweet potato (Ipomoea batatas,Convolvulaceae) complex

• Genome size evolution and its relationship with pollen grain size has been investigated in sweet potato (Ipomoea batatas), an economically important crop which is closely related to diploid and tetraploid species, assessing the nuclear DNA content of 22 accessions from five Ipomoea species, ten sweet potato varieties and two outgroup taxa.
• Nuclear DNA amounts were determined using flow cytometry. Pollen grains were studied using scanning and transmission electron microscopy.
• 2C DNA content of hexaploid I. batatas ranged between 3.12–3.29 pg; the mean monoploid genome size being 0.539 pg (527 Mbp), similar to the related diploid accessions. In tetraploid species I. trifida and I. tabascana, 2C DNA content was, respectively, 2.07 and 2.03 pg. In the diploid species closely related to sweet potato e.g. I. ×leucantha, I. tiliacea, I. trifida and I. triloba, 2C DNA content was 1.01–1.12 pg. However, two diploid outgroup species, I. setosa and I. purpurea, were clearly different from the other diploid species, with 2C of 1.47–1.49 pg; they also have larger chromosomes. The I. batatas genome presents 60.0% AT bases.
• DNA content and ploidy level were positively correlated within this complex. In I. batatas and the more closely related species I. trifida, the genome size and ploidy levels were correlated with pollen size. Our results allow us to propose alternative or complementary hypotheses to that currently proposed for the formation of hexaploid Ipomoea batatas.

     

发根农杆菌介导药用甘薯西蒙1号的遗传转化

用发根农杆菌A4分别感染药用甘薯西蒙1号的叶片、茎切段、叶柄等外植体,诱导出毛状根,并对毛状根进行了离体培养.采用L9(34)正交设计法优化甘薯西蒙1号的毛状根诱导条件;PCR扩增检测转化毛状根;用高效液相色谱仪检测了毛状根中咖啡酸的含量.结果表明:转化中茎切段是最合适的外植体,最佳感染时间20 min,共培养最佳时间为2天;PCR扩增检测表明发根农杆菌Ri质粒的T-DNA片段已整合进植物的基因组中;经高效液相色谱仪证实毛状根中含有咖啡酸,含量为0.03792 mg/g.     

Identification of the New Pathogen (Stemphylium lycopersici) Causing Leaf Spot on Pepino (Solanum muricatum)

Pepino (Solanum muricatum var. pepino) plants were found affected by an extensive leaf spot caused by plant pathogenic fungi during a survey in the Cameron highlands, Pahang state, Malaysia. Symptomatic leaf samples were collected from infected pepino plants and cultivated on PDA medium, and the pathogen was isolated and purified; then, consequently, all isolates were identified as Stemphylium lycopersici on the basis of their cultural and morphological characteristics and combined sequences of the internal transcribed spacer (ITS) and glyceraldehyde‐3‐phosphate dehydrogenase (gpd) regions. A pathogenicity assay on detached leaves further confirmed that S. lycopersici causes leaf spot disease. To the best of our knowledge, this is the first report of S. lycopersici causing leaf spot on pepino in Malaysia and worldwide.     

转昆虫抗冻蛋白基因增强甘薯抗冻能力

为明确昆虫抗冻蛋白基因转入甘薯(Ipomoea batatas)后是否能提升其抗冻能力,进而为培育甘薯抗冻育种材料奠定基础,将黄粉虫(Tenebrio molitor)抗冻蛋白基因TmAFP导入植物基因表达质粒,经农杆菌介导的遗传转化获得抗冻甘薯新材料。以甘薯品种Huachano为受体材料建立甘薯植株高效再生体系,并采用不同成分的体细胞胚成熟培养基培养胚性悬浮细胞。胚性愈伤组织对除草剂的敏感性测试结果表明,转基因阳性植株筛选的最适培养基为MS+0.2 mg·L–12,4-D+0.8 mg·L^–1 GAP+100 mg·L^–1 Carb。将表达质粒分别转化Huachano后共获得7个胚性愈伤团并最终获得42株再生抗性植株,其中转pSUIBEV3-AFP有23个株系,转pCAMBIA-AFP有19个株系,经PCR、Southern杂交和RT-PCR检测后证实TmAFP基因已整合至甘薯基因组中并获得表达。将转基因甘薯及对照植株在–1℃下处理15小时后转移至室温,结果表明,转基因甘薯植株的抗冻能力显著提升。     

Effects of Sweet Potato Feathery Mottle Virus and Sweet Potato Chlorotic Stunt Virus on the Yield of SweetPotato in Uganda

Sweet potato feathery mottle virus (SPFMV) and Sweet potato chlorotic stunt virus (SPCSV) are the most common viruses infecting sweetpotato in Uganda. Field plots planted with graft inoculated plants of virus‐free cultivars Beauregard, Dimbuka, Ejumula, Kabode and NASPOT 1 were used to assess the effect of SPFMV and SPCSV on yield and quality of sweetpotatoes in two agro‐ecologies. SPFMV spreads rapidly to control plots at Makerere University Agricultural Research Institute Kabanyolo (MUARIK), and these plots had similar yields to those singly infected with SPFMV but at the National Semi Arid Resource Research Institute (NaSARRI) where SPFMV spreads slowly, plots infected with SPFMV yielded 40% less than the control. Recovery from SPFMV appeared to be more frequent at NaSARRI than at MUARIK. Infection by SPCSV alone resulted in yield losses of 14–52%, while mixed infections of SPFMV+SPCSV resulted in yield losses in both locations of 60–95% depending on the cultivar. SPCSV and mixed infections of SPFMV+SPCSV also reduced the number of roots formed as well as the diameter of the roots, resulting in a greater length to diameter ratio compared to the healthy control. This study, therefore, confirms that both SPFMV and SPCSV, both singly and when mixed, can reduce yield, the extent depending on the cultivar. To mitigate the effect of these viruses, farmers should use clean planting materials of resistant varieties.     

First Report of Leaf Spot Disease in Angelica dahurica Caused by Phoma bellidis in China

Angelica dahurica is a perennial herb belonging to the Umbelliferae family that has been used as a valuable traditional Chinese medicine. In August 2013, leaf spot disease in A. dahurica was first observed at the Medicinal Herb Garden in Shenyang Agricultural University, Shenyang, Liaoning, China. Morphological assessment, evaluation of culture characteristics, internal transcribed spacer and beta‐tubulin sequence analysis and pathogenicity identified the causal agent as Phoma bellidis. To our knowledge, this is the first report of leaf spot disease in A. dahurica that was caused by P. bellidis.     

Survival of Sclerotia of Rhizoctonia solani AG3PT and Effect of Soil‐Borne Inoculum Density on Disease Development on Potato

Sclerotia produced by a single isolate of Rhizoctonia solani AG3PT were buried in small plot experiments to investigate the effects of sclerotial production method, soil type and burial depth on sclerotial viability in field soil. The factor with the greatest effect on sclerotial viability, defined as the percentage of sclerotia germinating on agar following retrieval, in all experiments was the duration of burial. After 18 months, on average across all experiments, 20% of retrieved sclerotia were viable. A comparison between sclerotia produced in vitro on malt yeast extract agar and in vivo using micropropagated tubers in field soil found no significant differences between the two production methods on sclerotial viability. Burial in field soil at 20‐cm depth was found to significantly reduce sclerotial viability to 50% compared to 60% at 5 cm. In two pot experiments, amending the growing medium and soil with increasing inoculum densities of R. solani was found to increase stem number, stem canker and black scurf severity regardless of whether this soil‐borne inoculum was derived from mycelium or sclerotia. Black scurf incidence and severity were assessed 30–32 days posthaulm destruction and found to be similar for a range of sclerotial soil‐borne inoculum densities (1.0 × 10^?1 g/kg d.w. soil to 6 × 10^?3 g/kg d.w. soil). The significance of these findings in relation to pathogen survival, detection in soil and disease development is discussed.     

Anthracnose Caused by Colletotrichum horii on Sweet Persimmon in Korea: Dissemination of Conidia and Disease Development

Anthracnose disease caused by Colletotrichum horii (C. gloeosporioides), results in considerable economic damage to sweet persimmon in southern Korea yearly. This study deals with the life cycle of the pathogen in terms of seasonal fluctuations of spore dispersal and the development of disease based on field surveys, spore potential and fungal isolation. Anthracnose disease was observed first on twigs in the last week of May and reached an incidence of 1.2%. Subsequently, the disease increased rapidly and reached an incidence of 86% by the end of July. Infection of fruits started in mid‐June (2.8%) and increased gradually to 64.4% by the end of July. In severely infected orchards, 46.2% of diseased fruits were dropped. The pathogen began releasing conidia in the first week of May and continued until the end of September. The maximum release of spores was observed in mid‐July. To determine the optimal use of chemicals for control of anthracnose, the following spray programme was evaluated. Spraying two or three times resulted in 89.4 and 93% control, respectively, whereas spraying more than four times led to 100% control. In comparison, the disease rate of unsprayed trees was 89.8%. To control anthracnose effectively, it is recommended to take steps to eliminate inoculum sources in sweet persimmon orchards before spraying chemicals.     

Occurrence of Passalora Leaf Spot Caused by Passalora ampelopsidis on Virginia Creeper in Korea

A previously unknown leaf spot disease was observed on Virginia creeper (Parthenocissus quinquefolia) vines in Seoul, Korea. Affected plants exhibited leaf spots and discoloration, resulting in leaf blight and premature defoliation. The causal agent of the disease was identified as Passalora ampelopsidis by comparing morphological characteristics and analyzing the internal transcribed spacer region of the rDNA. Pathogenicity tests were conducted twice with the same results, fulfilling Koch's postulates. Passalora leaf spot on Virginia creeper vines was first recorded in the United States in 1878. Recent reports of Passalora leaf spots on Virginia creeper vines in 2005 in Germany, in 2012 in China and now in Korea indicate the prevalence of the disease.     

Sweet potato (Ipomoea batatas) trypsin inhibitors expressed in transgenic tobacco plants confer resistance against Spodoptera litura

A sweet potato (Ipomoea batatas cv. Tainong 57) trypsin inhibitor gene was introduced into tobacco plants (Nicotiana tabaccum cv. W38) by Agrobacterium tumefaciens– mediated transformation. From 30 independent transformants, three lines with high level of expression were further analyzed. The trypsin inhibitor gene, under control of the 35S CaMV promoter, led to the production of the trypsin inhibitor proteins up to 0.2% of the total protein. In insecticidal bioassays of transgenic tobacco plants, larval, growth of Spodoptera litura (F.), the tobacco cutworm, was severely retarded as compared to their growth on control plants. This observation implied that expression of sweet potato trypsin inhibitor can provide an efficient method for crop protection. Received: 29 July 1996 / Revision received: 15 November 1996 / Accepted: 8 December 1996     

Resistance to sweet potato virus disease (SPVD) in wild East African Ipomoea

Sweet potato virus disease (SPVD), the most harmful disease of sweet potatoes in East Africa, is caused by mixed infection with sweet potato feathery mottle potyvirus (SPFMV) and sweet potato chlorotic stunt crinivirus (SPCSV). Wild Ipomoea spp. native to East Africa (J cairica, I. hildebrandtii, I. involucra and J wightii) were graft-inoculated with SPVD-affected sweet potato scions. Inoculated plants were monitored for symptom development and tested for SPFMV and SPCSV by grafting to the indicator plant J setosa, and by enzyme-linked immunosorbent assay (ELISA). Virus-free scions of sweet potato cv. Jersey were grafted onto these wild Ipomoea spp. in the field, and scions collected 3 wk later were rooted in the greenhouse and tested for viruses using serological tests and bioassays. In all virus tests, J cairica and J involucra were not infected with either SPFMV or SPCSV. J wightii was infected with SPFMV, but not SPCSV, in the field and following experimental inoculation; J hildebrandtii was infected with SPCSV, but not SPFMV, following experimental inoculation. These data provide the first evidence of East African wild Ipomoea germplasm resistant to the viruses causing SPVD.     

Perpetuation of Cherry Leaf Spot Disease in Ornamental Cherry

Cherry leaf spot disease, caused by Blumeriella jaapii (Rehm) Arx., is an increasing concern to nursery producers of ornamental cherry in the south‐eastern United States. Spores were trapped starting in late March before symptoms were observed in the field, which indicates that leaf debris from diseased trees are an important source of primary inoculum. Previously infected trees of six cultivars (‘Kwanzan’, ‘Yoshino’, ‘Okami’, ‘Snowgoose’, ‘Autumnalis’ and ‘Akebono’), which were overwintered in a controlled environment protected from airborne spores, developed disease symptoms in late spring, indicating that dormant buds may also be a source of primary inoculum. Because ornamental cherry trees are propagated by budding and cuttings, disease management should incorporate cultural practices that focus on propagation from disease‐free trees and fungicide applications beginning at petal drop to protect emerging leaves.     

甘薯根腐病菌侵染对甘薯内源激素水平的影响

甘薯根腐病病原菌[Fusarium solani(Mart.)Sacc.f.sp.batatas McClure,简称FSB]侵染及其培养液滤液处理高敏感性甘薯品种‘胜利百号’后,引起甘薯叶片、茎尖和根部组织内源ABA含量大幅升高。其中在根部出现最早,但茎尖中积累浓度最高。侵染后甘薯叶片、茎尖和根部组织内源GA1/3含量显著低于对照。甘薯组培苗经FSB培养滤液处理9h后,ABA含量显著上升,处理15h,ABA含量呈下降趋势,而GA1/3含量在101和102稀释液处理15h(103稀释液处理12h)时出现显著上升。这些结果有助于解释甘薯根腐病株矮小不产生藤蔓,并在秋季大量现蕾开花的生理现象。     

Zonate Leaf Spot of Prunus mandshurica Caused by Hinomyces pruni in Korea

Zonate leaf spots and severe defoliation were observed on Manchurian apricot (Prunus mandshurica) growing in a humid location in Korea from 2011 through 2013. The main symptoms included greyish green to brownish grey and zonate leaf spots without border lines, which mostly led to premature defoliation. The morphological characteristics of the causal agent were consistent with Hinomyces pruni. Identification was supported by analysing the sequence of the internal transcribed spacer region of ribosomal DNA from an isolate. The pathogenicity of the isolate was confirmed by artificial inoculation. This is the first report of zonate leaf spot caused by H. pruni on Manchurian apricot globally as well as in Korea.     

Identification and Characterization of Paraphoma chrysanthemicola Causing Leaf Spot Disease on Atractylodes japonica in China

Atractylodes japonica is a perennial herb in Compositae family, which is used for stomach disorders as a traditional Chinese medicine (Guo et al. 2006 ). In 2013, a leaf spot disease was first observed on plants of A. japonica in a production field of Fushun County, Liaoning Province, China. The disease had a speckled appearance initially. Lesions with grey–white centre and brown margin gradually developed and enlarged. Eventually, infections usually caused yellowing of the leaves and premature defoliation. The causal agent of infection on plants was identified as Paraphoma chrysanthemicola based on morphological and cultural characteristics, pathogenicity tests and phylogenetic analysis. To our knowledge, this is the first report of a leaf spot disease on A. japonica caused by P. chrysanthemicola in China.     

First Report of Leaf Blight Disease Caused by Fusarium verticillioides (Teleomorph Gibberella moniliformis) in Kentucky Bluegrass

Leaf blight symptom caused by Gibberella moniliformis was observed in the seedlings of Kentucky bluegrass (cv. Perfection) in Jinju, Korea, in January 2013. The symptoms first appeared as brown to yellow lesions on leaves, which later became white and bleached. The pathogen was isolated from turfgrass leaves and identified as G. moniliformis by using the internal transcribed spacer (ITS)1 and ITS4 sequences of rRNA. G. moniliformis was confirmed by a pathogenicity test, and the causal agent was reisolated from infected turfgrass.     

Occurrence of Grey Leaf Spot of Sedum erythrostictum Caused by Cercospora cf. pseudokalanchoes in China

Sedum erythrostictum is a perennial herb in the Crassulaceae family, which is a traditional Chinese medicine used for the treatment of hepatitis, dysentery, herpes zoster and swellings. In 2014, a grey leaf spot disease causing significant damage to plants of S. erythrostictum occurred in the Medicinal Herb Garden of Jilin Agricultural University, Jilin Province, China. The fungus mainly infected the leaves. The necrotic lesions on the leaves were circular or elliptical, amphigenous, greyish brown to brown, slightly concave and surrounded by a dark brown distinct margin. The causal agent from symptomatic tissues was identified as Cercospora cf. pseudokalanchoes based on the symptoms, morphological characteristics, molecular identifications and pathogenicity tests. To our knowledge, this is the first formal report of grey leaf spot of S. erythrostictum caused by C. cf. pseudokalanchoes in China.     

Determination of Acylated Anthocyanin in Human Urine after Ingesting a Purple-Fleshed Sweet Potato Beverage with Various Contents of Anthocyanin by LC-ESI-MS/MS

Eighty-seven healthy volunteers ingested a purple-fleshed sweet potato beverage with various contents of anthocyanin (beverage A; 22.1 mg/250 ml, B; 107.8, C; 84.9). An acylated anthocyanin, peonidin 3-caffeoylsophoroside-5-glucoside, was detected in the urine 2 h after ingestion. The concentrations were 15.1±2.2 μg/l of urine (mean±SEM), 46.6±5.3, and 53.3±2.2 for beverages A, B, and C respectively.