A field screening method to evaluate rice breeding lines for resistance to the hoja blanca virus

A method is presented for rearing large colonies of viruliferous Sogatodes oryzicola, vector of the rice hoja blanca virus (RHBV). These colonies were used for field screening up to 10 000 rice breeding lines per season for resistance to RHBV. Uniform infection of check varieties in the field indicated that the method was adequate. Field release of vectors when plants were 14 days old resulted in satisfactory disease incidence, after 21 days, to distinguish lines segregating for resistance from lines uniformly resistant or susceptible. Various sources of resistance identified earlier continued to be resistant under the screening conditions. Progeny of lines identified as non-segregating resistant continued as non-segregating resistant. Resistant plants from lines segregating for resistance produced progeny lines that were segregating and non-segregating. Ratios of resistant to susceptible plants in F₁ progeny of three-way crosses were consistent with earlier observations that RHBV resistance is a dominant character. The susceptibility of the commercial checks indicates that rice production in RHBV areas of tropical Latin America continues to be at risk from the virus. Virus-resistant commercial cultivars resulting from this method should be available in 2 years.     

Evaluation of methods of screening strawberry cultivars for resistance to crown rot caused by Phytophthora cactorum

Four methods were evaluated in measuring resistance of strawberry cultivars to crown rot caused by Phytophthora cactorum. Meristem propagated plants grown in vitro were inoculated with mycelial discs. Four to five days after inoculation, it was possible to distinguish between cultivars with large differences in susceptibility to the disease. Ten days later, all plants were totally necrotic making it impossible to distinguish between cultivars. When detached leaves were inoculated by inoculating a plug of mycelium into the petiole, disease symptoms developed more slowly in resistant cultivars, but leaf age greatly affected the rate of symptom development. When plug plants (not cold stored) were lightly wounded in the rhizome with a scalpel and inoculated with either zoospores or mycelium, differences in disease development between cultivars were mainly as would be expected from previous information on susceptibility, but both age and size of plants influenced the rate of disease development. Unwounded, inoculated plants did not develop symptoms. When cold‐stored plug plants were either unwounded or lightly wounded with a scalpel in the rhizome and inoculated with zoospores, the relative rates of disease development consistently reflected the susceptibility to crown rot. At the time of final assessment, disease was much more severe in wounded plants, but the relative susceptibility of cultivars was not affected by the wounding.     

The interactions of potato cyst nematodes and Verticillium dahliae on early and maincrop potato cultivars

The effects of potato cyst nematodes (Globodera rostochiensis and G. pallida) on the growth of early and later maturing potato cultivars, with or without the H, resistance gene, were compared in pot experiments and the effects of introducing Verticillium dahliae into the system were studied. The influence of the nematodes on root function and structural integrity was inferred from their effects on calcium and water uptake. Early maturing cultivars were less tolerant of nematode attack than later maturing ones, and those with the H₁ resistance gene were more tolerant of G. rostochiensis than G. pallida. When V. dahliae was introduced, Verticillium wilt symptoms appeared much earlier on the early than the maincrop cultivars. Also, Maris Anchor (first early) died before Maris Peer (second early) and death of both when infected by nematodes and fungus was much earlier than uninfected control plants or plants infected with G. rostochiensis only. Maris Peer infected by both organisms snowed symptoms much more severe than when infected by either organism alone, whereas Maris Anchor was almost as severely affected by V. dahliae alone as both organisms together. Pentland Javelin (first early) developed Verticillium wilt symptoms several weeks later than other cultivars and the addition of nematodes failed to increase the severity of the symptoms. These differences in cultivar susceptibility to the combination of nematodes and fungus may be due to the nematodes causing different degrees of trauma in the roots of the three cultivars: measuring calcium uptake per unit water uptake showed that nematodes dramatically increase calcium uptake but that the proportional change is least in Pentland Javelin. If this is a reflection of less damage by nematodes to the endodermis of this cultivar, it may explain the greater resistance of its roots to penetration by V. dahliae.     

Identification of RFLP markers for resistance to wheat spindle streak mosaic bymovirus (WSSMV) disease.

Wheat spindle streak mosaic bymovirus (WSSMV) causes an economically important disease of winter wheat in Europe and North America. Artificial inoculation with this virus to identify resistant wheat genotypes is difficult. This study was conducted to identify restriction fragment length polymorphism (RFLP) markers associated with resistance to this disease. A population, consisting of 104 F5 recombinant inbred lines from a cross between hexaploid Triticum aestivum cultivars 'Geneva' (resistant) and 'Augusta' (susceptible), was evaluated for WSSMV symptoms under field conditions for four years. Two linked markers on the long arm of chromosome 2D, Xbcd1095 and Xcdo373, were determined to be associated with WSSMV resistance by bulked segregant analysis of the 10 most resistant and 10 most susceptible lines. Marker Xcdo373 accounted for 79% and Xbcd1095 for 73% of the phenotypic variation. Our results suggest that resistance to WSSMV in this population is qualitative in nature and is controlled by few genes. These markers should be useful in the development of wheat cultivars resistant to WSSMV and perhaps also to wheat yellow mosaic bymovirus (WYMV).     

Resistance to Fusarium head Blight and Deoxynivalenol Accumulation in Chinese Barley

Fusarium head blight (FHB) caused by Fusarium graminearum Schwabe is a devastating barley disease world-wide, causing significant yield losses and contaminating cereal products with mycotoxins. Barley grain contaminated with deoxynivalenol (DON) is associated with gushing and may be rejected by the malting and brewing industry. Genetically inherited resistance is the most effective option for the control of the disease. A total of 266 barley cultivars and breeding lines originating from China were evaluated for FHB resistance and concentration of DON in grain. Plants were inoculated with isolates of F. graminearum under field conditions by injecting conidia into a single spikelet of each spike. FHB symptoms were evaluated by visual inspection, and DON content was analysed by HPLC. Significant differences in FHB ratings and DON levels were observed among cultivars. Visual symptoms of FHB varied from 4.88 to 71.75% of infected spikelets 21 days after inoculation and from 7.86 to 113.33 area under the disease progress curve units (AUDPC). Twenty-seven lines were more resistant to FHB than the control resistant cultivar Zhedar 2 and with fewer than 12% infected spikelets. Twenty-one of the above lines originated from the area in the mid to low valley of Yangtze River, where FHB epidemics are frequent. DON levels ranged from 0.05 to 24.39 mg/kg among the tested barley lines. Correlation coefficients were significant between FHB symptom ratings and DON levels. However, there was no significant correlation between symptom rating and plant height and no significant correlation between symptom rating and heading date.     

Transgenic banana expressing Pflp gene confers enhanced resistance to Xanthomonas wilt disease

Banana Xanthomonas wilt (BXW), caused by Xanthomonas campestris pv. musacearum, is one of the most important diseases of banana (Musa sp.) and currently considered as the biggest threat to banana production in Great Lakes region of East and Central Africa. The pathogen is highly contagious and its spread has endangered the livelihood of millions of farmers who rely on banana for food and income. The development of disease resistant banana cultivars remains a high priority since farmers are reluctant to employ labor-intensive disease control measures and there is no host plant resistance among banana cultivars. In this study, we demonstrate that BXW can be efficiently controlled using transgenic technology. Transgenic bananas expressing the plant ferredoxin-like protein (Pflp) gene under the regulation of the constitutive CaMV35S promoter were generated using embryogenic cell suspensions of banana. These transgenic lines were characterized by molecular analysis. After challenge with X. campestris pv. musacearum transgenic lines showed high resistance. About 67% of transgenic lines evaluated were completely resistant to BXW. These transgenic lines did not show any disease symptoms after artificial inoculation of in vitro plants under laboratory conditions as well as potted plants in the screen-house, whereas non-transgenic control plants showed severe symptoms resulting in complete wilting. This study confirms that expression of the Pflp gene in banana results in enhanced resistance to BXW. This transgenic technology can provide a timely solution to the BXW pandemic.     

THE USE OF TETRACHLORONITROBENZENE ISOMERS ON LETTUCE

Field experiments were carried out from 1951 to 1955 on winter lettuce plants grown in boxes under glass and in the open, and on commercial crops in unheated glasshouses. Each of the three isomers of tetrachloronitrobenzene, applied as 5% dusts at 1/4 and 1/2 oz. per square yard of bed, gave significant protection against Botrytis disease, but the 2:3:4:5 isomer was inferior to the other two isomers and they all delayed the hearting of the crop. The 2:3:4:5 isomer at both rates of application and the 2:3:5:6 and 2:3:4:6 isomers at 1/2 oz./sq.yd. delayed hearting of a commercial crop by more than 14 days and are not recommended at these rates. Local overdoses of fungicide should be avoided.     

54份CIMMYT小麦材料抗白粉病分析

选用来自我国不同地区的20个白粉病菌毒性菌株,对54个CIMMYT小麦品种(系)进行抗病性分析.结果表明:(1)34个品种(系)含有抗病基因,以Pm8基因出现频率最高,有15个品种(系)携带该基因;(2)参试主效基因中,Pm1、Pm3e、Pm5、Pm6和Pm7基因已丧失对我国白粉菌的抗性,Pm16和Pm20基因的抗性最强;(3)50个1B/1R易位系品种(系)中31个含有抗病基因,48%的抗病1B/1R易位系可检测到Pm8基因.根据田间成株期病程曲线下面积(AUDPC)聚类分析结果,可将54份材料分为高抗、中抗、中感和高感4类,7个品种(系)不含任何主效抗病基因而田间表现中到高的抗性,是典型慢病性品种.     

Evaluation of Rice Cultivars under Greenhouse Conditions for Adult-Plant Resistance to Pyricularia oryzae

Six rice cultivars showing various types of resistance or susceptibility to Pyricularia oryzae in the field were compared under controlled environmental conditions. The resistance of the cultivars with adult-plant resistance was race-specificat early growth stage. On all cultivars tested, blast infection became increasingly reduced on either leaves of adult plants or older leaves, as observed in rice plants of different leaf stages infected with different individual races. Their increase inhost resistance was marked by an apparent transition in infection type and reduced blast severity depending on leaf age and developmental stage of plants. The ranking of disease severity of the adult-plantresistant cultivars to different races was constant during plant development, whereas that of the susceptible cultivars was differential. It is suggested that different degrees of resistance in rice cultivars may exist in seedlings and be consistently maintained during plant development, probably becoming more distinct as rice plants mature.     

Resistance reactions of Cucumis melo to inoculation with Pseudoperonospora cubensis

Reactions of several cultivars and breeding lines of melons to artificial inoculation at the seedling stage with a local isolate of Pseudoperonospora cubensis were compared under growth chamber and field conditions. Four different reactions were distinguishable: highly susceptible, susceptible, partially resistant and resistant. The differences in the reactions were more clear-cut under growth chamber conditions. Selection for resistance could be carried out at the seedling stage using sporangia production and lesion size, shape, colour and number as criteria. Under growth chamber conditions, plants of resistant lines had minimal sporangia production and circular or angular lesions 1–2 cm in diameter which became necrotic within 5 days after inoculation.     

Virus Movement Protein Gene Mediated Resistance Against Cucumber Mosaic Virus Infection

Tobacco (Nicotiana tabactum L. ) "NC89" plants were transformed with deletion mutant of cucumber mosaic virus (CMV) movement protein (MP) gene and full-length CMV MP gene, respectively. The transformed plants were analyzed with polymerase chain reaction (PCR), PCR-Southem, Southern and Western blots. R0 generation of the transgenic plants were inoculated with CMV. Five out of 10 lines of tobacco plants (BMPK) transformed with CMV MP deletion mutant gene showed high resistance to CMV infection and remained symptomless for up to 50 days post-inoculation. In contrast, tobacco plants (BMPR) transformed with full-length CMV MP gene did not show resistance to CMV infection. However, most of the infected full-length CMV MP gene transgenic plants recovered by showing none or very mild mosaic symptoms in 40 days post-inoculation. The results of R1 generation of the BMPK transgenic plants tested under field conditions showed that all 5 lines of transgenic plants could delay the virus disease development.     

Occurrence and physico-serological properties of potato virus S (PVS) in Iran

1818 collected samples of potato plant showing virus infection symptoms from 85 fields were tested for PVS infection using DAS-ELISA. Average of infection to this virus varied from 0 to 100%. Least infection was belonging to fields with new introduced varieties. On the other hand native and old introduced cultivars showed heavy infection. In field condition, PVS infected plants didn't show very obvious symptoms, so some infected plants may be missed in field sample collecting. The physical properties of 3 isolates, Avaj, Stanboly and Agria No 15 were determined. TIP 55-60 degrees C, DEP 10(-3) and Liv measured 3-4 days. Ouchterlony agar double diffusion test using SDS was useful for virus detection and precipitation lines didn't show any spur between isolates, although isolates differs slightly in symptomatology. SDS-Page and Western blotting methods used successfully for virus detection and determining and measuring viral protein components.     

A glasshouse screening method to assess resistance in onions to damage by onion fly (Delia antiqua) larvae

Factors influencing the resistance of onion (Allium cepa) cultivars to onion fly (Delia antiqua) damage were investigated. These included different environments, methods of plant raising and egg inoculation, and plant size. A glasshouse compartment maintained at 14 to 37 °C was as satisfactory for assessing the resistance as a controlled environment. The basic test unit comprised 10 onion seedlings raised in a 400 × 50 × 50 mm deep plastic trough and inoculated with 50 onion fly eggs from a laboratory culture. Resistance was assessed by counting wilted plants every two or three days for 21 days after egg inoculation. The reciprocal of number of days for each plant to wilt (wilting rate) was an appropriate variate for statistical analysis, adjusted for the length of the longest leaf at the time of inoculation. The method enables cultivars or breeding lines of onion species to be reliably screened for resistance to larvae of this pest in a glasshouse so that only the most promising need be tested in the field.     

Replication of tomato yellow leaf curl virus (TYLCV) DNA in agroinoculated leaf discs from selected tomato genotypes

The leaf disc agroinoculation system was applied to study tomato yellow leaf curl virus (TYLCV) replication in explants from susceptible and resistant tomato genotypes. This system was also evaluated as a potential selection tool in breeding programmes for TYLCV resistance. Leaf discs were incubated with a head-to-tail dimer of the TYLCV genome cloned into the Ti plasmid ofAgrobacterium tumefaciens. In leaf discs from susceptible cultivars (Lycopersicon esculentum) TYLCV single-stranded genomic DNA and its double-stranded DNA forms appeared within 2–5 days after inoculation. Whiteflies (Bemisia tabaci) efficiently transmitted the TYLCV disease to tomato test plants following acquisition feeding on agroinoculated tomato leaf discs. This indicates that infective viral particles have been produced and have reached the phloem cells of the explant where they can be acquired by the insects. Plants regenerated from agroinfected leaf discs of sensitive tomato cultivars exhibited disease symptoms and contained TYLCV DNA concentrations similar to those present in field-infected tomato plants, indicating that TYLCV can move out from the leaf disc into the regenerating plant. Leaf discs from accessions of the wild tomato species immune to whitefly-mediated inoculation,L. chilense LA1969 andL. hirsutum LA1777, did not support TYLCV DNA replication. Leaf discs from plants tolerant to TYLCV issued from breeding programmes behaved like leaf discs from susceptible cultivars.The Hebrew University of Jerusalem, Faculty of Agriculture, Department of Field and Vegetable Crops     

Comparison of Resistance to Cotton Leaf Curl Disease (Multan/Burewala) in Gossypium hirsutum L. Varieties and Breeding Lines

Twenty‐four cotton varieties and advance breeding lines were evaluated for their resistance to cotton leaf curl disease (CLCuD; Multan/Burewala) under natural field and in glasshouse conditions for two consecutive years. Resistance was based on symptom expression and disease severity index. All the cotton genotypes exposed to the vector whitefly in the field and artificially infected by grafting manifested a high level of resistance against CLCuD (Multan) with the exception of genotype NIAB‐999 that was moderately resistant. All the test varieties/breeding lines were highly susceptible to CLCuD (Burewala) both in the field and the glasshouse. However, substantial differences were noted between genotypes for disease index under field conditions. Graft inoculation studies showed that all genotypes inoculated with CLCuD (Burewala) developed disease within 9–13 days whereas those graft‐inoculated with CLCuD (Multan) developed symptoms from 15 to 22 days after grafting. Severe reduction occurred in plant morphology, fibre and yield parameters of cotton variety NIAB‐111 following inoculation with CLCuD (Burewala) as compared with CLCuD (Multan).     

山东省12个主栽小麦品种(系)抗叶锈性分析

本研究旨在明确山东省12个小麦主栽品种(系)抗叶锈性及抗叶锈基因,为小麦品种推广与合理布局、叶锈病防治及抗病育种提供依据。利用2015年采自山东省的5个小麦叶锈菌流行小种的混合小种对这些材料进行苗期抗性鉴定,然后选用15个小麦叶锈菌生理小种对这些品种(系)进行苗期基因推导,并利用与24个小麦抗叶锈基因紧密连锁(或共分离)的30个分子标记对其进行抗叶锈基因分子检测。结果显示,山东省12个主栽小麦品种(系)苗期对该省2015年的5个小麦叶锈菌混合流行小种均表现高度感病。通过基因推导与分子检测发现,济南17含有Lr16,矮抗58和山农20含有Lr26,其余济麦系列、烟农系列、良星系列等9个品种(系)均未检测到所供试标记片段。此外,本研究还对山东省3个非主栽品种进行了检测,结果发现,中麦175含有抗叶锈基因Lr1和Lr37,含有成株抗性基因;皖麦38只检测到Lr26,济麦20未检测到所供试标记片段。综合以上结果,山东省主栽小麦品种(系)所含抗叶锈基因丰富度较低,尤其不含有对我国小麦叶锈菌流行小种有效的抗锈基因,应该引起高度重视,今后育种工作应注重引入其他抗叶锈基因,提高抗叶锈性。     

Selection for Fusarium wilt disease resistance from regenerants derived from leaf callus of strawberry

Resistant lines of strawberry to the fungal wilt disease caused by Fusarium oxysporum f. sp. fragariae were selected strawberry plants regenerated from leaf-derived callus tissues. Regenerants were transplanted to a field heavily infested with this pathogen, and normally growing plants were selected as the putative resistant lines. Daughter plants produced vegetatively through runner formation of the lines were similarly tested in the pathogen-infested field over an additional three generations. Finally, two resistant lines were obtained from a total of 1,225 regenerants. The stable propagation of disease resistance in these lines was confirmed by directly inoculating the daughter plants with the pathogen and planting in a pathogen-infested soil. All of the control plants were efficiently infected and died within one month. The isolated plant lines grew and developed runners even after direct inoculation and produced daughter plants in this soil. Thus, the present study demonstrated the existence of somaclonal variation for disease resistance against a soil-borne fngal pathogen.