The origin of rhizomania resistance in sugar beet

In the last 35 years, breeding has greatly reduced the damages caused by rhizomania in sugar beet crops. After the first encouraging results using the Alba genotypes, the cultivar Rizor represented a substantial step forward and has given good yield improvement in diseased fields in many parts of the world. The original variety and subsequent improved versions continued to offer good performances for about a decade, after which it was surpassed by other hybrids derived in part from the Rizor itself. Further progress in terms of sugar production became possible in 1986, when the Holly monogerm lines were released in USA and Europe. In spite of the incomplete information about the genealogy of the first resistant materials,many evidences and the molecular analyses on the different genotypes suggest a possible common progenitor and lineage. The resistant cultivars have kept the yield at an adequate level, allowing cultivation to continue in countries where the disease has reached epidemic proportions. The case of rhizomania resistance in sugar beet can therefore be considered as one of the most important achievements in plant breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

A greenhouse test for screening sugar-beet (Beta vulgaris) for resistance to beet necrotic yellow vein virus (BNYVV)

Small differences in activity between batches of purified beet necrotic yellow vein virus (BNYVV) were observed in ELISA. A four-parameter modelled dose-response curve of purified BNYVV was used for the conversion of ELISA values to virus concentrations. Seedlings of the susceptible cultivar Regina and the partially resistant cultivars Nymphe and Rima were tested for resistance to BNYVV in a mixture of sand and infested soil. Plants were grown in a green-house with low nutrient supply and at temperatures below the optimum of both the vectorPolymyxa betae and BNYVV. Root systems were small and consisted mainly of lateral roots. Significant differences in average virus concentrations were found between cultivars, both when using the complete root systems and when using either the top or the bottom part of the root systems. Average virus concentrations in Regina were always significantly higher than in Rima and higher than in Nymphe on all occasions except one (P<0.05). Differences between Nymphe and Rima were less evident. Variation between plants was greatest within Rima. The test described in this paper can be used for the discrimination of different cultivars and for the identification of individual plants with resistance to BNYVV.     

Development of a recombinant antibody ELISA test for the detection of Polymyxa betae and its use in resistance screening

An ELISA test was developed for the quantitative detection of the obligate parasite Polymyxa betae , the vector of Beet necrotic yellow vein virus (BNYVV), in infected sugarbeet roots. The test used monoclonal and polyclonal antibodies raised to a recombinantly expressed glutathione-S-transferase (GST) from P. betae . A close correlation was found between the number of P. betae zoospores in serially diluted suspensions and absorbance values in the ELISA test. Time-course studies of plants grown in naturally infested soils in controlled environment tests demonstrated the value of the ELISA test in screening for P. betae resistance. In preliminary tests, P. betae -resistant accessions of the wild sea beet ( Beta vulgaris ssp. maritima ), which might be used to restrict the transmission of BNYVV, were identified.     

Partial nucleotide sequence analysis of Turkish isolates of Beet necrotic yellow vein virus (BNYVV) RNA-3

A survey was carried out to detect Beet necrotic yellow vein virus (BNYVV) in soil samples using RT-PCR and bait plant techniques from the sugar beet production area of Tokat, Turkey in 2001. More than 80% of the soil samples analyzed were found to be contaminated with the virus. The partial nucleotide sequence of cDNA corresponding to RNA-3 of BNYVV isolates were analyzed for six different regions of Tokat province. All isolates were assigned to type A strains based on RFLP analysis and DNA sequences. Sequence comparison revealed differences at amino acid positions 35, 68, 71 and 179 of the P25 coding region amongst Turkish isolates. Additionally, all Turkish isolates were compared with Japanese, French, Kazakh, Italian and Belgian isolates.     

Effect of sugar beet cultivars with different levels of resistance to beet necrotic yellow vein virus on transmission of virus byPolymyxa betae

The effect of resistance of sugar beet cultivars to beet necrotic yellow vein virus (BNYVV) on virus content of resting spore clusters of the vectorPolymyxa betae was studied in controlled environments and in naturally infested fields. The total number of resting spore clusters formed in roots of a partially resistant and a susceptible cultivar did not differ when assessed 6 and 12 weeks after inoculation with viruliferous resting spores. Transmission experiments showed that in partially resistant plants, having a low virus content in the roots, the population of resting spores formed was less viruliferous than that in susceptible plants with a high virus content. Consequently, growing a resistant cultivar can be expected to delay the build-up of virus inoculum in soil.In a trial field sampled in 1991, the inoculum potential of BNYVV (most probable number of viruliferousP. betae propagules) in soil was lower after growing a partially resistant cultivar than after growing a susceptible one. On the other hand, in four sites sampled in 1990, inoculum potential in soil was hardly increased by growing sugar beet and was not significantly affected by the cultivar grown.     

甜菜抗丛根病种质创新与分子生物技术

甜菜坏死黄脉病毒（BNYVV）及其新变异株P型和IV-A型仍对甜菜生产造成重大损失。选育抗病品种是防治该病的唯一有效措施。用ELISA和实时定量逆转录PCR技术测定BNYVV和P．betae可对杂交后代分离群体或转基因后代中的抗病个体进行选择。AFLP、KAPD和PCR等分子标记的建立可对抗病基因位点Rz1（Holly）、Rz2（WB42）、Rz3（WB41）和Rz4（C50,R22）进行辅助选择。在分离植物抗病基因方面,已获得含Rz1的BAC克隆和候选基因。转基因甜菜含重复倒置的BNYVV复制酶基因、RNA-2转运蛋白P15突变基因或CP基因显著提高了抗病性。国际合作可有效地利用资源,加快抗病种质创新。     

QTL mapping of BNYVV resistance from the WB258 source in sugar beet

Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV), is an important sugar‐beet disease worldwide. The aim of this study was to investigate and map BNYVV resistance from the WB258 source. Presence of a major resistance gene (termed Rz5) was indicated by ～1 : 1 segregation of virus levels in a population of 170 individuals derived from WB258. The maximum logarithm of odds (LOD) position for Rz5, identified by interval mapping, explained ～81% of the phenotypic variance. Rz5 was mapped alongside the previously identified Rz4 from the R36 source. Rz4 and Rz5 were located to 6.9 cM and 6.0 cM regions, respectively, on chromosome III by interval mapping. These locations corresponded well to the position of the widely deployed Rz1 gene, suggesting that the three genes could represent an allelic series. Physical mapping is probably required to confirm this however. BNYVV resistance genes from novel sources are potentially important in breeding for more durable resistance either as separate genes or as BNYVV race‐specific alleles.     

甜菜丛根病研究进展

甜菜丛根病是一种发生在世界主要糖甜菜种植地区的土传病害,在缺乏有效控制的情况下会引起甜菜严重的产量损失。从4个方面叙述了近些年来与甜菜丛根病研究的相关进展：①甜菜坏死黄脉病毒(Beet necrotic yellow vein virus,BNYVV)的基因组结构;②BNYVV的分类及与其他甜菜土传病毒的关系;③丛根病的检测方法;④丛根病的控制策略。最后对甜菜丛根病未来的研究发展方向进行了展望。     

Resistance against rhizomania disease via RNA silencing in sugar beet

Rhizomania is one of the most damaging and widely spread diseases in major sugar beet growing regions of the world. The causal agent, beet necrotic yellow vein virus (BNYVV), is transmitted via the fungus Polymyxa betae, which retains it in the field for years. In this study, an RNA silencing mechanism was employed to induce resistance against rhizomania using intron‐hairpin RNA (ihpRNA) constructs. These constructs were based on sequences of the BNYVV 5′‐untranslated region of RNA‐2 or the flanking sequence encoding P21 coat protein, with different lengths and orientations. Both transient and stable transformation methods produced effective resistance against rhizomania correlated with the transgene presence. Among the constructs, those generating ihpRNA structures with small intronic loops produced the highest frequencies of resistant events. The inheritance of transgenes and resistance was confirmed over generations in stably transformed plants.     

Breeding for resistance to rhizomania in sugar-beet (Beta vulgaris L.)

In breeding for resistance to rhizomania, breeding material was routinely examined for resistance to the rhizomania virus over several years. This study was performed in order to investigate the value of virological examination of sugarbeet breeding material in breeding for resistance to the rhizomania virus. Tests were conducted with varieties susceptible or partially resistant to rhizomania, as well as with several new and more-resistant hybrids. A test on young plants showed that all genotypes contained beet necrotic yellow-vein virus (BNYVV), but that the virus content differed widely. Plants of two commercially used partially resistant varieties had, on average, only half the virus content of susceptible varieties in their roots, and plants of eight new hybrids still in the process of registration contained less than one third of that amount. There was a significant negative correlation between the quantity of BNYVV in young plant roots and final yield under disease conditions in the field. Opportunities for further improvement in the level of the resistance to BNYVV in sugarbeet by means of conventional breeding programmes are discussed.