Identification and Mapping of Two New Genes Conferring Resistance to Powdery Mildew from Aegilops tauschii （Coss,） Schmal

Two powdery mildew resistance genes were Identified from Aegilops tauschll accessions Y201 and Y212 and mapped using two different F2 populations derived from the crosses between susceptible accession Y2272 and Y201, and susceptible accession Y2263 and Y212. Genetic analysis of resistance to powdery mildew Indicated that the resistance of Y201 was controlled by a single dominant gene, whereas the resistance of Y212 was controlled by a single recessive gene. We have temporarily designated these genes as PmY201 and PmY212, respectively. By bulk segregation analysis, six mlcrosatelllte markers Including Xgwm174, cfd26, cfd57, cfdl02, Xgwm583 and Xgwm639 were found to be linked to PraY201 with genetic distances of 5.2, 7.7, 9.6, 12.5, 20.2 and 22.1 cM, respectively. Five SSR markers, including cfd57, Xgwm182, cfd7, cfd102, and cfd12, were found to be linked to PmY212 with distances of 5.6, 7.2, 11.5, 14.7, and 18.5 cM, respectively. According to the locations of the linked markers, the two resistance genes were located In the 5DL region. Based on the chromosomal locations and the resistance patterns of the two genes, we propose that PmY201 and PmY212 are two novel powdery mildew resistance genes, and are suitable for marker-assisted selection.     

Gene Network and Database for Genes of Wheat’s Resistance to Pathogenic Fungi

An overview describing a gene network that controls the formation of plant responses to diseases caused by pathogenic fungi (http://wwwmgs.bionet.nsc.ru/mgs/gnw/genenet//viewer/Plant%20fungus%20pathogen.html) is presented. The gene network represents the coordinated interactions of genes, proteins, and regulatory molecules, including integrated defense mechanisms that prevent the development of infection, localize the lesion, and minimize damage. The gene network was reconstructed on the basis of literature data, and the elements of the gene network were associated with the records of the PGR database (Pathogenesis-Related Genes, http://srs6.bionet.nsc.ru/srs6bin/cgi-bin/wgetz?-page+top+-newId), where information on plant genes resistant to pathogenic fungi is accumulated. Reconstruction of the gene network allows us to formalize, visualize, and systematize possible mechanisms for the response of plant cells to fungal infection, which may be useful for the planning of experiments and interpretation of experimental data in this field of science.     

Characterization of DNA methylation variations during fruit development and ripening of Vitis vinifera (cv. ‘Fujiminori’)

Physiology and Molecular Biology of Plants - The fruit is the most important economical organ in the grape; accordingly, to investigate the grapevine genomic methylation landscape and examine its...     

Transgenic plantlets of ‘Chancellor’ grapevine (Vitis sp.) from biolistic transformation of embryogenic cell suspensions

Transgenic plantlets of Chancellor grapevine (Vitis L. complex interspecific hybrid) were produced via biolistic transformation. Embryogenic cell suspensions were bombarded with 1 m tungsten particles coated with pBI426 which encodes a fusion peptide between -glucuronidase (GUS) and neomycin phosphotransferase II (NPTII). The fusion peptide is under the control of a double 35S Cauliflower Mosaic Virus promoter and a leader sequence from Alfalfa Mosaic Virus. The cells were placed on kanamycin-containing media (10, 25 or 50 mg/l) 2 d after bombardment. Activated charcoal reduced cell browning. Embryos were first observed on selective media 14–29 weeks after bombardment. More than 1600 clusters of embryos were germinated and/or assayed for GUS. Of 621 embryos assayed for GUS expression, 182 (29.3%) were positive. PCR confirmed the presence of the NPTII gene in all 5 GUS-positive and 2 GUS-negative (bombarded) embryos tested. In germination experiments, 15% of the embryo clusters produced at least one plant with normal shoot growth. Of 164 normal plants assayed for GUS expression, 37 (22.6%) were positive. The NPTII gene was amplified by PCR in 1 (of 1) GUS-positive and 4 (of 5) GUS-negative bombarded plants, but not in non-bombarded control plants. Southern blotting confirmed integration of the NPTII gene in all 3 of the GUS and PCR-NPTII positive plants tested. Biolistics is an efficient method for transformation of Chancellor and should be applicable to other important grape cultivars.Abbreviations AC activated charcoal - GUS -glucuronidase - 2,4-D 2,4-dichlorophenoxyacetic acid - BA 6-benzylaminopurine - NAA -naphthalene acetic acid - TDZ thidiazuron - NPTII neomycin phosphotransferase II - Km kanamycin - MS Murashige and Skoog (1962) medium - WPM Woody Plant Medium of Lloyd and McCown (1980)     

Changes in Water Uptake in Light and Darkness of the Barley Cultivar ‘Professor Schiemann’ Infected with Powdery Mildew

In the investigated barley cultivar resistant toErysiphe graminis f. sp.hordei Marchal a temporary decrease in water uptake may be observed during the first days following infection in light. No marked differences could be detected in darkness. Starting from the stage of formation of fructification organs the ratio between water uptake in light and in darkness in the resistant cultivar is similar to that of the control, while the opposite is true in the susceptible cultivar where the ratio markedly decreases with the development of powdery mildew. The resistant cultivar responds more rapidly to the pathogen by changes in water uptake than the susceptible one.     

Transcriptomic Analysis Revealed Hormone-Related and Receptor-Like Kinase Genes Involved in Wound Healing of ‘Duli’ and its Resistance to Valsa Pyri

Plant Molecular Biology Reporter - Valsa canker is a destructive fungal disease that results in a serious loss of production. The pathogen Valsa pyri (Vp) usually infiltrates the bark and xylem via...     

Effect of phyllotactic position and cultural treatments toward successful direct shoot organogenesis in dwarf ‘Pixie’ grapevine (Vitis vinifera L.)

In Vitis spp. where somatic embryogenesis-based regeneration predominates, an efficient, reproducible and robust method of direct shoot organogenesis from leaf explant material has been established in the dwarf wine grape ‘Pixie’ (Vitis vinifera). This regeneration system was achieved by testing the response of leaf material in two stages of development, and pre-conditioning the explant material in dark conditions and/or in liquid media prior to excising from the plant and placing it on solidified media. The pre-excision treatments included (1) a dark period of 24 h, with no regeneration medium; (2) soaking in regeneration medium followed by a dark period of 24 h; (3) a dark period of 24 h followed by soaking in liquid VRM (Vitis Regeneration Medium); (4) vacuum infiltration in liquid VRM followed by a dark period of 24 h; and (5) a control of no pre-conditioning treatment. Excised leaves from pre-treated intact plants in vitro significantly increased the frequency of shoot organogenesis. The most responsive explant material consisted of young semi-translucent apical leaves varying in size from 3 to 8 mm in length. The most successful combinations of factors contributing to shoot organogenesis involved the solely dark-exposed apical leaves or the soaking in VRM followed by a dark period. These results are expected to facilitate Vitis-related research in genetics, functional genomics, physiology, and other fields.     

Effects of Shading on the Synthesis of Volatile Organic Compounds in ‘Marselan’ Grape Berries (Vitis vinifera L.)

Journal of Plant Growth Regulation - Light is a vital environmental factor that can affect the synthesis of volatile organic compounds (VOCs) in grape berries. However, the mechanism through which...     

Plant regeneration via somatic embryogenesis from solid and suspension cultures of Vitis vinifera L. cv. ‘Manicure Finger’

Vitis vinifera L. cv. ‘Manicure Finger’ is one of the major table grape varieties in China. To provide a strong foundation for genetic transformation with potential for crop improvement, we undertook plant regeneration via somatic embryogenesis. Anthers and gynoecia were harvested from immature flowers and used as explants to induce embryogenic calli. Explants cultured in MS1 medium (based on Murashige and Skoog basal salts), supplemented with 4.5-μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.4-μM 6-benzylaminopurine (6-BA) showed the highest rates of embryogenic callus induction (3.7%?±?1.3% for anthers and 4.8%?±?2.5% for gynoecia). After several months, somatic embryos were produced from embryogenic calli cultured in plant growth regulator-free MS2 medium (with reduced sucrose). Somatic embryos (SE) at the cotyledonary stage were isolated and cultured on three different media (MS2, MS3, or B) for conversion into plantlets, the efficiency of which ranged from 63.9%?±?4.8% to 83.9%?±?8.4%. After 1 mo of in vitro culture, 80% of plants with at least six leaves were successfully transplanted into soil. SE was repeatedly induced from previously induced somatic embryos for up to 1.5 yr. Using embryogenic calli as starting material, suspension cultures containing embryogenic cell aggregates were also established in liquid MS medium supplemented with 4.5-μM 2,4-D. The embryogenic cell aggregates continued to proliferate without differentiating for successive subculture cycles. After transfer to 2,4-D-free liquid medium for 4 wk, an average of 63.7%?±?9.0% mature SEs were produced per 20 mL of liquid medium. More than 40% of somatic embryos at cotyledonary stage, derived from the suspension cultures, successfully germinated into plants using solid medium.     

Evaluation of Construct and Criterion Validity for the ‘Liverpool Osteoarthritis in Dogs’ (LOAD) Clinical Metrology Instrument and Comparison to Two Other Instruments

Objective

To test the ‘Liverpool Osteoarthritis in Dogs’ (LOAD) questionnaire for construct and criterion validity, and to similarly test the Helsinki Chronic Pain Index (HCPI) and the Canine Brief Pain Inventory (CBPI).

Design

Prospective Study.

Animals

222 dogs with osteoarthritis.

Procedure

Osteoarthritis was diagnosed in a cohort of dogs on the basis of clinical history and orthopedic examination. Force-platform analysis was performed and a “symmetry index” for peak vertical force (PVF) was calculated. Owners completed LOAD, CBPI and HCPI instruments. As a test of construct validity, inter-instrument correlations were calculated. As a test of criterion validity, the correlations between instrument scores and PVF symmetry scores were calculated. Additionally, internal consistency of all instruments was calculated and compared to those previously reported. Factor analysis is reported for the first time for LOAD, and is compared to that previously reported for CBPI and HCPI.

Results

Significant moderate correlations were found between all instruments, implying construct validity for all instruments. Significant weak correlations were found between LOAD scores and PVF symmetry index, and between CBPI scores and PVF symmetry index.

Conclusion and Clinical Relevance

LOAD is an owner-completed clinical metrology instrument that can be recommended for the measurement of canine osteoarthritis. It is convenient to use, validated and, as demonstrated here for the first time, has a correlation with force-platform data.     

Assaying for Pollen Drift from Transgenic ‘Rainbow’ to Nontransgenic ‘Kapoho’ Papaya under Commercial and Experimental Field Conditions in Hawaii

In 1992, papaya ringspot virus (PRSV) was discovered in the Puna district of Hawaii island where 95% of the state of Hawaii’s papaya was being grown. By 1998 production in Puna had decreased 50% from 1992 levels. A PRSV-resistant transgenic papaya ‘Rainbow’ containing the coat protein gene of PRSV was released commercially in Hawaii in 1998, and saved the papaya industry from further devastation. In the ensuing years since the release of the transgenic papaya, a number of farmers grew hermaphrodite nontransgenic ‘Kapoho’ papaya in close proximity to plantings of hermaphrodite transgenic ‘Rainbow’ papaya. These plantings provided a unique opportunity to assay for transgenic-pollen drift under commercial conditions. Between 2004 and 2010, assays for the GUS (beta-glucuronidase) transgene in embryos were done to study transgenic-pollen drift in commercial ‘Kapoho’ plantings and in replicated field plots. Very low pollen drift (0.8%) was detected in fruit of ‘Kapoho’ trees in the border row of one plantation when 90 embryos were assayed per fruit, while no pollen drift was detected in four other commercial plantings in which eight embryos were tested per fruit. Pollen drift averaged 1.3% of tested embryos in field plots where individual hermaphrodite ‘Kapoho’ trees were adjacent to two or four ‘Rainbow’ trees. In contrast, 67.4% of tested embryos were GUS positive in similarly located female ‘Kapoho’ trees. The very low transgene flow to close-by ‘Kapoho’ plantings is likely due to the fact that hermaphrodite trees are used commercially in Hawaii and that these trees are largely self-pollinated before the stigma is exposed to external pollen.     

Genetic analysis of two wheat cultivars, ‘Sonalika’ and ‘WL 711’ for reaction to leaf rust (Puccinia recondita)

Summary Genetic analysis for leaf rust reaction of two widely adapted cultivars, Sonalika and WL 711, has been done using 21 near isogenic Lr lines and rust culture IL004 — avirulent on the two cultivars and all the Lr lines used. The segregation pattern in the F₂ generation indicated the presence of a recessive gene in Sonalika and of a dominant gene in WL 711. These genes in cultivars Sonalika and WL 711 have been identified as Lr 11 and Lr 13, respectively. Gene Lr 13 is no longer effective in WL 711 but it continues to give field resistance in the backgrounds of Chris, Prelude and Thatcher. There has been no significant change in the virulence spectrum of the leaf rust pathogen in India with the release of WL 711. High susceptibility of WL 711 seems to be due to the evolution of more aggressive forms of the pathogen to this cultivar. The gene Lr 11, which behaves as a recessive in Sonalika, was effective against leaf rust when this cultivar was released. The high susceptibility of Sonalika is probably due to an increase in the frequency of race 77 virulent on Lr 11. Lr 11 has shown a dominance reversal in the background of Sonalika. Present results suggest that interaction of resistance genes with the background genotype must be studied for their effective use in breeding programme.     

Pandemic Serotypes of Vibrio cholerae Isolated from Ships’ Ballast Tanks and Coastal Waters: Assessment of Antibiotic Resistance and Virulence Genes (tcpA and ctxA)

There is concern that ships’ ballasting operations may disseminate Vibrio cholerae to ports throughout the world. Given evidence that the bacterium is indeed transported by ships, we isolated pandemic serotypes O1 and O139 from ballast tanks and characterized them with respect to antibiotic resistance and virulence genes ctxA and tcpA. We carried out concurrent studies with V. cholerae isolated from coastal waters. Of 284 isolates, 30 were serotype O1 and 59 were serotype O139. These serotypes were overrepresented in ballast tanks relative to the coastal waters sampled. All locations, whether coastal waters or ballast tanks, yielded samples from which serotype O1, O139, or both were isolated. There were three groups among the 62 isolates for which antibiotic characterization was conclusive: those exhibiting β-lactamase activity and resistance to at least one of the 12 antibiotics tested; those negative for β-lactamase but having antibiotic resistance; those negative for β-lactamase and registering no antibiotic resistance. When present, antibiotic resistance in nearly all cases was to ampicillin; resistance to multiple antibiotics was uncommon. PCR assays revealed that none of the isolates contained the ctxA gene and only two isolates, one O139 and one O1, contained the tcpA gene; both isolates originated from ballast water. These results support the bacteriological regulations proposed by the International Maritime Association for discharged ballast water.     

Eriophyoid mite damage in Vitis vinifera (grapevine) in Australia: Calepitrimerus vitis and Colomerus vitis (Acari: Eriophyidae) as the common cause of the widespread ‘Restricted Spring Growth’ syndrome

Leaf and shoot distortions and retarded shoot growth in Vitis vinifera L. prevalent in Australian vineyards in early spring, were investigated in replicated field experiments over 3 yrs. Leaf distortion and retarded shoot growth were identified as damage due to feeding of extremely high populations of over-wintered deutogynes of Calepitrimerus vitis (Nalepa) (grape rust mite). This damage was hitherto known in Australia as Restricted Spring Growth (RSG), a syndrome comprising several growth abnormality symptoms, none with a clearly identified cause or a successful treatment. A successful treatment against C. vitis was used to selectively eliminate RSG, while C. vitis numbers were recorded using a validated trapping technique; intercepting deutogynes migrating from winter shelters in the wooden vine structure, to emerging green tissues. Severe leaf distortion was associated with >400 C. vitis deutogynes per spur, while >1000 per spur had the added effect of severely retarding shoot growth. A 43.0–47.2% shoot length reduction was recorded for Cabernet Sauvignon, 27.1–32.8% for Sauvignon Blanc, when 4–6 leaves were separated. Symptoms were most prominent up to 8–9 separated leaves, however 24.7–30.4% shoot length reduction was still evident at flowering, and 12.8% circa fruit set. C. vitis effect on vine fruitfulness, and yield parameters at fruit set, were also studied. Once successfully treated to prevent C. vitis damage, poor bud burst remained evident in some vineyards. Surveys of unburst buds from such vineyards revealed presence of Colomerus vitis (Pagenstecher) (grape bud mite). When Col. vitis numbers in unburst buds reached 100–500 per bud, apical meristems of primary, and commonly also secondary buds were dead, preventing bud burst. The remaining living scale tissue was distinctly scarred. Bud and associated shoot damage were documented. Retarded shoot growth and leaf distortion, previously attributed to RSG, are misdiagnosed C. vitis spring feeding damage. Clustered high infestations of Col. vitis can cause bud-axis necrosis, bud burst failure, shoots with short basal internodes, and short, thin, zigzagged shoots with absent fruit clusters; all previously considered RSG.This revised version was published online in May 2005 with a corrected cover date.     

Recombinant expression,purification, and characterization of polyphenol oxidase 2 (VvPPO2) from “Shine Muscat” (Vitis labruscana Bailey × Vitis vinifera L.)

Polyphenol oxidases (PPOs) catalyze browning reactions in various plant organs, therefore controlling the reactions is important for the food industry. PPOs have been assumed to be involved in skin browning of white grape cultivars; however, the molecular mechanism underlying PPO-mediated browning process remains elusive. We have recently identified a new PPO gene named VvPPO2 from “Shine Muscat” (Vitis labruscana Bailey × V. vinifera L.), and have shown that the gene is transcribed at a higher level than the previously identified VvPPO1 in browning, physiologically disordered berry skins at the maturation stage. In this study, we expressed VvPPO2 in Escherichia coli and, using the purified preparation, revealed unique physicochemical characteristics of the enzyme. Our study opens up a way to not only understand the berry skin browning process but also to elucidate the enzymatic maturation process of grape PPOs.     

Synthesis and Evaluation of Novel Oligodeoxynucleotides Containing 3′-C-(3-Benzoyloxypropyl)thymidine and Bicyclo Nucleoside Derivatives

Abstract

The stereoselective synthesis of 3′-C-Allyluridine derivative 2 has been accomplished. This nucleoside was used as a key synthon for the synthesis of oligodeoxynucleotides containing 3′-C-(3-benzoyloxypropyl)thymidine (X) or bicyclo nucleoside (Y+Z) monomers. Preliminary thermal experiments are reported.