Acquisition of potato leafroll virus byMyzus persicae from secondarily-infected potato plants of different genotypes

Summary The acquisition of potato leafroll virus (PLRV) byMyzus persicae nymphs from the top leaves of potato plants was studied throughout a growing season in relation to the antigen titre in those leaves and the feeding behaviour of the aphid. Secondarily-infected plants of eight potato genotypes with different levels of field resistance served as virus sources. Early in the growing season, plants were efficient sources for virus acquisition. The amount of viral antigen detected inM. persicae nymphs fed on the top leaves was strongly correlated with the titres of viral antigen in these leaves. Virus acquisition from the top leaves of older potato plants was markedly impaired and could not be correlated with their virus titre. With increasing age of the potato plants and the development of virus symptoms, the virus titre in the leaves declined and the initial weak correlation between the virus titre and field resistance ratings disappeared. Thus, screening secondarily-infected potato plants for field resistance to PLRV based on the concentration of viral antigen in leaves or in aphids fed on them should be avoided later in the growing season. The feeding rate ofM. persicae, measured by the number of honeydew droplets excreted, did not account for the reduced uptake of virus from older plants since it was not influenced by the age of the plant. Throughout the growing season, the feeding rate ofM. persicae nymphs on PLRV-infected plants was higher on genotypes with low levels of field resistance to PLRV than on genotypes with high ones.     

A method of assessing potato seedling progenies for resistance to the white potato cyst nematode

Summary The resistance expressed by progenies of potato seedlings to the white potato cyst nematode (Globodera pallida) has been assessed on bulk sowings of seedlings and on individual plants raised from tubers, and a positive correlation between the seedling and plant tests has been established. The use of such a seedling test as an addition to other methods of screening is discussed.     

Enhanced <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">tuberosi</Emphasis> Resistance in Transgenic Potato Expressing a Rice GLP Superoxide Dismutase Gene

Germin like proteins (GLPs) are a large group of related and ubiquitous plant proteins which are considered to be involved in different processes important for plant development and defense. Multiple functional copies of this gene family have been reported in a number of species (wheat, barley, rice, soybean mosses and liverwort), and their role is being evaluated by gene regulation studies and transgenic approaches. To analyze the role of a rice (Oryza sativa) root expressed germin like protein1 OsRGLP1, for its antifungal activity, transgenic potato plants were developed. These transgenic potato plants were molecularly characterized and biologically assessed after inoculation with Fusarium oxysporum f. sp. tuberosi. Functional analysis showed high accumulation of H₂O₂, increased Superoxide Dismutase (SOD) activity and no oxalate oxidase activity (OxO) in transgenics in comparison to nontransformed control. This increased SOD activity, resistance to heat and sensitivity to H₂O₂ suggest it is a Fe-like SOD. OsRGLP1 expression in potato plants exhibited enhanced resistance in comparison to nontransformed wild type plants suggesting its role in providing protection against Fusarium oxysporum f. sp. tuberosi through elevated SOD level. Overall, results suggest that OsRGLP1 is a candidate for the engineering of potato plants with increased fungal tolerance however, the greater height and tuber number was observed. This phenotype associated with the resistance needs to be evaluated to determine if this is a positive or negative feature.     

Biological factors influencing the transmission of potato leafroll virus by different aphid species

Summary Macrosiphum euphorbiae, collected in the field from potato plants infected with potato leafroll virus (PLRV), transmitted the virus to fewer potato plants in a field trial than did laboratory-rearedMyzus persicae. In the laboratory,M. persicae was the only efficient vector of PLRV fromPhysalis floridana seedlings, potato sprouts or excised leaves toP. floridana. Two clones ofM. euphorbiae and one clone ofAulacorthum solani transmitted PLRV from infected potato plants toNicotiana clevelandii as effeciently asM. persicae but a clone ofAphis gossypii was an inefficient PLRV vector. An isolate of PLRV, whichM. persicae transmitted inefficiently from potato toN. clevelandii, was also transmitted inefficiently byM. euphorbiae andA. solani.     

TaCYP78A5基因过表达小麦的遗传和功能初步分析

为了验证小麦籽粒大小相关基因TaCYP78A5在小麦籽粒发育中的功能,对pINO启动子驱动的TaCYP78A5基因过表达的转基因小麦后代株系进行了鉴定,检测了T_0代植株目标基因拷贝数,定量分析了7个T_1代阳性植株的目标基因表达,并对其籽粒大小进行了统计。结果表明,利用Bar试纸条和目标基因特异PCR检测相结合的方法对21株转基因T_0代再生苗进行检测,共鉴定出14个阳性植株,除2个植株的目标基因拷贝数为3和1个植株为7外,其余11个T_0代转基因植株目标基因插入拷贝数均为1~2个,其中有6个单拷贝植株。与野生型相比,7个T_1代阳性植株目标基因表达量均极显著增加,粒厚和粒宽均有不同程度增加,粒重极显著增加。     

Cisgenesis,a New Tool for Traditional Plant Breeding,Should be Exempted from the Regulation on Genetically Modified Organisms in a Step by Step Approach

Modern potato breeding requires over 100,000 seedlings per new variety. Main reasons are (1) the increasing number of traits that have to be combined in this tetraploid vegetatively propagated crop, and (2) an increasing number of traits (e.g., resistance to biotic stress) originates from wild species. Pre-breeding by introgression or induced translocation is an expensive way of transferring single traits (such as R-genes, coding for resistance to biotic stress) to the cultivated plant. The most important obstacle is simultaneous transfer of undesired neighbouring alien alleles as linkage drag. Stacking several genes from different wild sources is increasing this linkage drag problem tremendously. Biotechnology has enabled transformation of alien genes into the plant. Initially, transgenes were originating mainly from microorganisms, viruses or non-crossable plant species, or they were chimeric. Moreover, selection markers coding for antibiotic resistance or herbicide resistance were needed. Transgenes are a new gene source for plant breeding and, therefore, additional regulations like the EU Directive 2001/18/EC were developed. Because of a strong opposition against genetic modification of plants in Europe, the application of this Directive is strict, very expensive, hampering the introduction of genetically modified (GM) crops and the use of this technology by small and medium-sized enterprises (SMEs). Currently, GM crops are almost the exclusive domain of multinationals. Meanwhile, not only transgenes but also natural genes from the plant species itself or from crossable plant species, called cisgenes, are available and the alien selection genes can be avoided in the end product. This opens the way for cisgenic crops without alien genes. The existing EU directive for GM organisms is not designed for this new development. The cisgenes belong to the existing breeders’ gene pool. The use of this classical gene pool has been regulated already in agreements regarding breeders’ rights. We are proposing a step by step approach starting with a crop and gene specific derogation and monitoring towards a general exemption of cisgenic plants from the Directive. Two examples, i.e. development of cisgenic potato for resistance to Phytophthora infestans and cisgenic apple for resistance to Venturia inaequalis are discussed shortly for illustration of the importance of cisgenesis as a new tool for traditional plant breeding. Cisgenesis is simplifying introgression and induced translocation breeding tremendously and is highly recommended for SMEs and developing countries.     

The expression of resistance to the white potato cyst nematode (Globodera pallida) by individual potato seedlings under different inoculation treatments

Summary The resistance toGlobodera pallida of individual potato seedlings from three progenies derived fromSolanum tuberosum ssp.andigena CPC 2802 was assessed using three inoculation treatments. Plants grown from tubers from the seedlings were used to reassess resistance. All the seedling test inoculation treatments over-estimated the proportion of clones classified as resistant compared to the results from tuber-grown plants. However, the mean relative resistance of progenies estimated by both tests showed good agreement, and it is suggested that seedlings are best used for assessing the resistance of progenies of crosses to identify promising combinations rather than individual clones.     

Detecting resistance to the root-knot nematodesMeloidogyne hapla andM. chitwoodi in potato and wildSolanum spp.

Summary Various methods of screening for resistance to root-knot nematodes were compared and evaluated. Seedling populations ofSolanum spp., grown in clay pots and plastic tubes with silver sand and inoculated with juveniles ofMeloidogyne chitwoodi andM. hapla, showed large differences in the number of egg masses on roots 7 weeks after inoculation. The differences were reproducible when re-testing was done with cuttings and plants from tubers. No resistance toMeloidogyne spp. was observed with ten potato cultivars when grown in clay pots, plastic tobes or closed containers. Plants from tubers in growth pouches developed a large two-dimensional root system, and after inoculation with juveniles the infection process could be observed over 8 weeks. A method of infecting potato tuber tissue withMeloidogyne is described, using tuber slices in Petri dishes as a potential screening test for tuber resistance.     

Resistance to <Emphasis Type="Italic">Potato virus Y</Emphasis> in a Multitrait Potato Breeding Scheme without Direct Selection in Each Generation

In an experimental breeding scheme to improve late blight (Phytophthora infestans) and white potato cyst nematode (Globodera pallida) resistance of tetraploid potato over three generations of crossing and selection, 15 clones survived the final selection, and these were derived from 15 great-grandparents. There was no direct selection for resistance to Potato virus Y (PVY), but 14 out of the 15 great-grandparents were resistant to PVY and three had extreme resistance. Thirteen of the 15 descendants had PVY resistance and one extreme resistance. This was within the range expected for a random (unselected) sample from the genotypes of the great-grandparents. Hence, we found no evidence for any positive or negative association between PVY resistance and the attributes selected. The conclusion is that laborious selection is not required in every generation when many parents have PVY resistance, including some with more than one copy of a PVY resistance gene or resistance at more than one locus. However, in the future, determining the major virus resistance genes present in potential parents in each generation using diagnostic molecular markers would prevent susceptible × susceptible crosses being made and maximise the number of resistant × resistant ones.     

Systemic induced resistance to late blight in potato by treatment with salicylic acid andPhytophthora cryptogea

Summary Tests for systemic induced resistance (SIR) to late blight were performed with different potato cultivars in field trials and in growth chamber experiments. The non-pathogenPhytophthora cryptogea (Pc) and salicylic acid (Sa) were used as inducer agents. In the field, plants were naturally infected byPhytophthora infestans while the indoor plants were inoculated. The degree of SIR obtained varied with the cultivar used and also with the type of inducer. For example in the field the susceptible cv. St. Cecilia had enhanced resistance with Pc whereas Sa caused increased susceptibility. Inbred resistant cvs acquired increased resistance after treatment with Sa. In a separate pilot experiment in a growth chamber, Sa at two different concentrations was injected into potato seed tubers before planting. The higher Sa concentration increased resistance in genetically resistant cultivars while the lower Sa concentration brought about increased susceptibility.     

Extreme resistance to infection by potato virus X in genotypes of wild tuber-bearingSolanum species

Summary Clones derived from thirty-one different accessions (nineteen of Argentine origin) belonging to eightSolanum species were screened for resistance to infection by potato virus X strain cp (PVX cp) by mechanical inoculation of plantlets that had been micropropagated in vitro. Estimates of PVX multiplication obtained by enzyme linked immunosorbent assay and slot blot nucleic acid hybridization allowed the identification of resistant clones derived from five accessions belonging toS. commersonii S. oplocense, S. sparsipilum andS. tuberosum andigena. Resistant genotypes supported PVX concentrations 5 to 15 times smaller than did the susceptible control cultivar Spunta. Graft inoculation test confirmed the presence of extreme resistance similar to that conferred by the ‘immunity’ gene X¹ (also called RX_act).     

Resistance to late blight (Phytophthora infestans (Mont.) de Bary) of potato plants regenerated fromin vitro selected calli

Summary Cells of seven potato cultivars were selectedin vitro with culture filtrate (CF) ofPhytophthora infestans (Mont.) de Bary. Regenerated plants were tested for late blight resistance. The aim of the study was to check the efficacy of CF in the selection of potatoes for resistance to late blight and to evaluate the effects of additive factors on general resistance. One selection cycle, applied to a cell culture system, selected cells resistant to toxic metabolites of CF.In vivo screening of clones regenerated from selected cells was done in two steps: on whole plants, assessing foliage late blight, and on detached leaves, assessing single factors of horizontal resistance. In general, the frequency of resistant variants selected with CF did not differ from that of resistant somaclonal variants. Nevertheless, high concentrations of CF in the growth medium seemed to induce an improvement of some partial resistance factors as compared with the source plants.     

普通小麦/山羊草属间杂交F_1的形态学及细胞遗传学研究

为了解普通小麦与山羊草属的杂交F1代的育性、细胞遗传学表现及育种利用潜力,以普通小麦7182和丰优1718为母本,分别以卵穗山羊草SY163和离果山羊草SY119为父本进行杂交,对杂交F1代的生育特性、染色体构型、田间条锈病抗性、F1自交和回交结实性等进行了分析。结果表明,F1代植株生长势较强,株型倾向母本,成熟期穗子易断,壳硬;杂交F1代高度不育,花粉可育率1.26%~3.54%,回交结实率2.08%~5.26%,仅普通小麦/卵穗山羊草自交结实,但结实率最高仅为0.25%。7182/SY163、7182/SY119、丰优1718/SY163、丰优1718/SY119杂交F1代花粉母细胞减数分裂中期I的染色体构型平均分别为31.11I+1.90II+0.03III、20.04I+7.45II+0.02III、30.08I+2.40II+0.04III、22.37I+6.30II+0.01III,说明普通小麦与2种山羊草杂交亲和性差,且可交配性主要由山羊草基因决定。田间条锈病抗性鉴定结果显示,F1代均对条锈菌混合小种高抗或免疫,这对于丰富小麦抗病基因资源有重要意义。     

Physiological effects of calonyctin on development and yield of potato

Summary Foliar sprays of calonyctin, a natural plant growth regulator extracted from the leaves ofCalonyction aculeatum (L.) House, at a concentration 0.05 activity units, increased the tuber yield of six potato (Solanum tuberosum L.) cultivars by 7–19% (mean 15%) during 1993 and 1994. Physiological differences between calonyctin-treated and untreated potato plants were usually detected 30–40 days after spraying them with calonyctin. The chlorophyll content of treated potato leaves increased, and the chlorophyll a:b ratio decreased. By 40 days after treatment the root oxidizability and soluble protein contents of different parts of the plants also increased. Their subsequent decline as the plants aged was inhibited by calonyctin. It is concluded that calonyctin delayed the senescence of potato plants and increased tuber growth at later stages of development.     

转TaSCL14基因小麦的遗传稳定性及农艺性状分析

为了获得农艺性状优良的转TaSCL14基因小麦品系,以普通小麦品种小偃39为受体获得的转TaSCL14基因小麦T1～T3代株(系)为材料,对其中TaSCL14基因的遗传稳定性和主要的农艺特性进行分析。结果表明,转TaSCL14基因小麦的T1和T2代植株的阳性率分别达到55.9%和85.6%,穗粒数、冬前分蘖与对照存在显著差异;T3代4个转基因株系3-4、3-7、3-8和3-11的阳性率达到90%以上,且TaSCL14基因在各株系中的表达水平都高于对照。与野生型相比,转基因小麦T3代部分株系的冬前分蘖、冬后分蘖、有效分蘖和小穗数均较野生型呈显著或极显著提高,但千粒重都较野生型有所降低,其中株系3-4和3-7降低幅度较大,分别达到显著和极显著水平。以上结果说明,TaSCL14基因的过表达能够影响转基因小麦的部分农艺特性,并且在株(系)间有差异。     

Testing the resistance to silver scurf in commercial potato cultivars under controlled environmental conditions

Summary Optimum storage conditions to identify resistance to silver scurf among commercially grown potato cultivars were determined in a series of experiments. Inoculation of field-grown tubers with a conidial suspension ofH. solani (concentration 10⁴ conidia per ml of water) and incubation at 15°C with 95% RH for 1 month followed by 2 months at 85% RH produced the greatest differences in disease severity between potato cultivars. Results with glasshouse-grown tubers were comparable with those from field-grown tubers, indicating that the test could be used to identify resistance in wild species of potato and to understand the inheritance of this resistance.     

农杆菌介导法将抗草甘膦基因转入玉米自交系

利用农杆菌介导法将抗草甘膦基因(ssu-G₂-aroA)转入优良玉米自交系R18-599和齐319,侵染后的愈伤组织转至潮霉素浓度为10 mg/L的筛选培养基上,继代筛选3轮,每轮3周;得到的抗性愈伤组织在含有潮霉素浓度为3 mg/L的分化培养基上进行分化,R18-599获得30株再生植株,齐319获得18株再生植株。经PCR鉴定后共得到7株转化抗性植株(4株R18-599、3株齐319)。经PCR-Southern及RT-PCR检测结果表明,ssu-G₂-aroA基因已稳定整合到了玉米基因组中,并在RNA水平上得到表达。     

An Outlook on the Localisation and Structure-Function Relationships of R Proteins in <Emphasis Type="Italic">Solanum</Emphasis>

The co-evolution of plants and plant-pathogens shaped a multi-layered defence system in plants, in which Resistance proteins (R proteins) play a significant role. A fundamental understanding of the functioning of these R proteins and their position in the broader defence system of the plant is essential. Sub-project 3 of the BIOEXPLOIT programme studies how R proteins are activated upon effector recognition and how recognition is conveyed in resistance signalling pathways, using the solanaceous R proteins Rx1 (from S. tuberosum spp. andigena; conferring extreme resistance against Potato Virus X), I-2 (from S. lycopersicon; mediating resistance to Fusarium oxysporum) and Mi-1.2 (from S. lycopersicon; conferring resistance to Meloidogyne incognita) as model systems. The results obtained in this project will serve as a model for other R proteins and will be translated to potential applications or alternative strategies for disease resistance. These include the modification of the recognition specificity of R proteins with the aim to obtain broad spectrum resistance to major pathogens in potato.