The transfer of the synaptic gene (sy-2) from 1EBN Solanum commersonii Dun. to 2EBN germplasm

Summary Crosses of a synaptic mutant (sy-2) of 2x(1EBN) S. commersonii Dun. with 2x(2EBN) S. chacoense Bitt. resulted in the production of 3x(2EBN) hybrids. Low levels of 2n egg production in these hybrids allowed subsequent crosses to be made with 4x(4EBN) cultivars, resulting in 5x(4EBN) S. commersonii-S. chacoense-Group Tuberosum hybrids. Seed set in these crosses averaged 0.22 seeds per fruit. Fifty-four unspotted seeds were produced in 1225 crosses of the 5x(4EBN) hybrids with 2x(2EBN) Group Phureja haploid extraction clones carrying dominant seed spot markers. These seeds yielded a total of 15 confirmed haploids having a diploid (2n=2x=24) or near diploid chromosome number. One haploid clone recovered displayed a synaptically abnormal phenotype similar to the original S. commersonii synaptic mutant. Preliminary test crosses of these haploids indicated that all haploids produced were 2EBN.This method for the transfer of alleles across EBN levels holds promise for the recovery of both qualitative and quantitative traits from 1EBN species. In these recovered, cross-compatible forms, clones with desired characteristics may be selected and easily incorporated into breeding programs.     

Assignment of Endosperm Balance Numbers to the tuber-bearing Solanums and their close non-tuber-bearing relatives

The Endosperm Balance Number (EBN) of over 80 species and subspecies of the tuber-bearing Solanums and their close non-tuber-bearing relatives representing 13 taxonomic series has been determined, with no species assigned to more than one EBN level. Among North American species, most diploids are 1EBN, most tetraploids are 2EBN and all hexaploids are 4EBN; however, among South American species most diploids are 2EBN, most tetraploids are 4EBN and again all hexaploids are 4EBN. Thus species may be isolated from others of the same ploidy level by EBN differences, e.g., 4×(2EBN) from 4×(4EBN), while other species differing in ploidy but having the same EBN may be intercrossed, e.g., 4×(2EBN) and 2×(2EBN). Chromosome doubling or 2n gametes can be used to make a lower EBN species compatible with a higher EBN species. These findings also explain the major crossing difficulties previously inherent in the use of North American species in potato improvement. They also have direct implications for potato improvement, barring the occurrence of other incompatibility barriers. Any 4×(4EBN) cultivar is endosperm compatible and thus will cross with 4×(4EBN) and 6×(4EBN) species. The 2×(2EBN) haploids of 4×(4EBN) cultivars likewise will hybridize with 2×(2EBN) and 4×(4EBN) species. All 2×(1EBN) species are crossable with 2×(2EBN) haploids through 2n gametes or chromosome doubling.Cooperative investigation of the U.S. Department of Agriculture, Agriculture Research Service, and the Wisconsin Experiment Station.     

The importance of Endosperm Balance Number in potato breeding and the evolution of tuber-bearing Solanum species

Summary Endosperm failure is considered the primary reason for the lack of success in intra-and interspecific crosses. The Endosperm Balance Number (EBN) hypothesis is a unifying concept for predicting endosperm function in intraspecific, interploidy, and interspecific crosses. In the EBN system, every species has an effective ploidy (EBN), which must be in a 2:1 maternal to paternal ratio in the endosperm for crosses to succeed. The knowledge of EBN is very useful in the transfer of genes from exotic germplasm, and in the development of new breeding schemes in potato. The paper describes the strategies for introducing 2x(1EBN), 2x(2EBN), 4x(2EBN) and 6x(4EBN) germplasm into the cultivated 4x(4EBN) potato gene pool. A new methodology for producing 4x(4EBN) and 2x(2EBN) chromosome addition lines is also discussed. EBN has evolutionary importance in the origin of tuber-bearing Solanums. The role of the EBN in the origin of diploid and polyploid potato species, and as a barrier for hybridization and speciation of sympatric species within the same ploidy level is demonstrated. The origin of 3x and 5x cultivated tuber-bearing Solanums may also be explained using the EBN concept. EBN has been reported to exist in other plant species: alfalfa, beans, blueberries, rice, soybeans, squashes, tomato, forage legumes, grasses, ornamentals and Datura stramonium. This indicates that EBN may have broad application and could be useful for germplasm transfer and breeding in other crop species.     

Male fertility and cytology of triploid hybrids between tetraploidSolanum commersonii (2n=4x=48, 2EBN) and Phureja-Tuberosum haploid hybrids (2n=2x=24, 2EBN)

Summary Solanum commersonii Dun. is a diploid (2n=2x=24, 1EBN) wild species of potential value for potato breeding. It is a reproductively isolated species and cannot be crossed with Tuberosum haploids (2n=2x=24, 2EBN) or other diploid 2EBNSolanum species. In order to overcome the EBN barriers, triploid hybrids were produced between Phureja-Tuberosum haploid hybrids, which form 2n pollen grains by parallel spindles, and tetraploidS. commersonii. Microsporogenesis analysis of the triploids indicated a trend towards low values of chromosome distribution at Anaphase I; lagging chromosomes were often observed as well. Despite these abnormalities, the percentage of stainable pollen was very high, ranging from 5.0% to 74.3%. A high variation in pollen grain diameter was also evident. Parallel and tripolar orientation of spindles at Metaphase II of microsporogenesis was a common feature of all the triploids analyzed, but dyads and triads were observed at a very low frequency. Therefore, also the frequency of 2n pollen was very low; the different size of stainable pollen appears to represent the ploidy levels which are possible according to the distribution of chromosomes in Anaphase I. The results obtained also suggest thatS. commersonii could have minor genes acting at the end of meiosis in such a way that, despite the presence of parallel/tripolar spindles, dyads/triads are not formed.Contribution no. 124 from the Research Center for Vegetable Breeding.     

Crossability relationships among Cajanus,atylosia and Rhynchosia species and detection of crossing barriers

Summary Crossability of two cultivars of Cajanus cajan, eight species of Atylosia and one of Rhynchosia was investigated. Of the 73 combinations attempted, success was achieved in 12 cases. C. cajan crossed successfully with A. albicans, A. cajanifolia, A. lineata, A. scarabaeoides, and A. trinervia. Within the genus Atylosia, A. lineata crossed with A. albicans and A. scarabaeoides, and A. scarabaeoides with A. sericea. Three species A. platycarpa, A. volubilis and R. rothii did not cross with any other one. In most of the unsuccessful combinations, although the pollen germinated on the receiving stigmas, the pollen tube growth was inhibited inside the stigma or in the stylar tissue.     

The breeding potential of wild potato species resistant to the potato tuber moth,Phthorimaea operculella (Zeller)

Summary The potato tuber moth, Phthorimaea operculella (Zeller) is an important pest of potatoes in the field and in stores in warm environments throughout the world. In this study genetic resistance to potato tuber moth was identified in clones of Solanum sparsipilum (coded MBN) originally developed for resistance to bacterial wilt and root-knot nematode. Resistance to this pest in S. sucrense and S. tarijense as well as S. sparsipilum was exploited in wide crosses with diploid and tetraploid cultivated potatoes, and haploids derived from S. tuberosum; hybrid progenies were produced. Crosses between resistant S. pinnatisectum or S. commersonii and cultivated potatoes failed completely, although S. commersonii did hybridise with two bridging species S. lignicaule and S. capsicibaccatum which are slightly compatible with cultivated potatoes. Resistance to potato tuber moth was transferred to all progenies except those in which S. tarijense was the resistant parent. The development of potatoes resistant to potato tuber moth is discussed in the context of population breeding for the lowland tropics.     

Introgression of the low browning trait from the wild Mexican species Solanum hjertingii into cultivated potato (S. tuberosum L.)

Internal discoloration of tubers resulting from impact damage (blackspot bruise) is a serious quality problem in potato production and utilization, reducing profits to growers and increasing costs for processors. Resistance to blackspot bruise has been identified in the wild species Solanum hjertingii and is therefore a potential germplasm resource for genetic resistance to this problem. A bridging cross between S. hjertingii and a cultivated diploid clone was used to produce a triploid hybrid population that exhibited very low tuber browning potential, indicating a dominant pattern of inheritance for this trait. The triploid progeny were subjected to in vitrochromosome doubling and the resulting hexaploid clones were screened for browning potential. A hexaploid clone selected for low browning was reciprocally crossed with cultivated S. tuberosum cultivars exhibiting high susceptibility to blackspot bruise. Tubers obtained from the seed progeny of these 4x-6x crosses (hereafter referred to as the BC₁ populations) were evaluated for browning potential and polyphenol oxidase (PPO) activity. Tubers from the BC₁ populations displayed a very low potential for melanin production, while PPO activity was quite variable. The low Pearson correlation coefficient (r² = 0.45), between browning potential and PPO activity suggests that the mechanism of blackspot bruise resistance derived from S. hjertingii cannot be explained simply as a reduction in the initial PPO activity. The expression of substantial resistance to browning and dominant expression pattern in these BC₁ progeny indicate that utilizing genetic elements derived from S. hjertingii provides a robust approach for developing blackspot bruise resistant potato varieties. This revised version was published online in August 2006 with corrections to the Cover Date.     

Tolerance to low temperatures and tuber soft rot in hybrids between Solanum commersonii and Solanum tuberosum obtained through manipulation of ploidy and endosperm balance number (EBN)

The objectives of this study were to evaluate the tolerance to low temperatures and tuber soft rot in hybrids between Solanum commersonii and Solanum tuberosum. The experimental materials consisted of F₁ triploid, BC₁ pentaploid‐near pentaploid and BC₂ tetraploid–near tetraploid hybrids. The F₁ triploids had a freezing tolerance and acclimatization capacity closest to S. commersonii. This indicated that the endosperm barriers which prevent the introgression of 1EBN S. commersonii into 4EBN S. tuberosum had been overcome. Indeed, the triploids produced 2n eggs, thus giving a compatible maternal to paternal EBN ratio in the hybrid endosperm generated by the 3x(2EBN) × 4x(3EBN) crosses. The tolerance to low temperatures of BC₁ and BC₂ hybrids was lower than that of the F₁. However, a number of genotypes were identified which were able to withstand temperatures down to ‐5°C. Some BC₂ hybrids were also tested for their tolerance to tuber soft rot, and some resistant hybrids were detected. A number of them combined the capacity for cold acclimatization with tolerance to tuber soft rot. These hybrids have an EBN of 4; they are fertile and have been used in backcrosses with 4EBN S. tuberosum.     

Interspecific hybridization between rice bean (Vigna umbellata) and its wild relative (V. minima): Fertility-sterility relationships

Summary The pre-and post-fertilization barriers in the interspecific crosses between Vigna umbellata and V. minima were investigated. In the reciprocal crosses (V. minima as the parent) the entry of pollen tubes into the ovary was delayed by about 4 h, and no seed set was observed. However, no pre-fertilization barriers were encountered in crosses involving V. minima as the parent and V. umbellata as the parent (normal cross). The delay/absence of divisions in the endosperm and the failure of embryo to divide were the post-fertilization barriers responsible for somatoplastic sterility in normal crosses which yielded a few hybrid seeds. The hybrid seeds showed poor germinability. The F₁ hybrids were intermediate between the parents in most morphological characters, and are completely sterile for pollen. Backcrossing of F₁ hybrid with both the parents did not restore fertility in the progenies. V. minima is considered as the tertiary gene pool of the rice bean.     

Tuber yield and quality of 4x-2x (FDR) potato progenies derived from the wild diploid species Solanum berthaultii and Solanum tarijense

Field experiments were carried out in order to evaluate 4x‐2x families derived from crosses between elite 4x potato cultivars and 2x Tuberosum‐Solanum tarijense and 2x Tuberosum‐Solanum berthaultii clones. Three traits were assessed: total tuber yield (TTY), haulm maturity (HM) and general tuber appearance (GTA). The degree of heterosis of these hybrid families was evaluated by comparison with the respective 2x and 4x parents as well as with seven 4x cultivars. The parental haploid species hybrids derived from S. berthaultii and S. tarijense combined two or more positive horticultural characteristics. Expressed as yield percentage of the 4x parents, the TTY of the families ranged from 53% to 246%. For TTY, the best 4x‐2x hybrid family ranked better than seven out of nine elite 4x cultivars. Some families had GTA scores in the range of the highly selected 4x cultivars. The families, however, were generally later maturing than the 4x parent group. Specific combining ability for TTY and GTA were the only two significant sources of variation observed in this genetic material. Parent‐offspring correlation coefficients were low for all traits, and indicated that parental performance would not be informative at either ploidy level. These results parallel previous investigations with distinct haploid species hybrids where a 4x‐2x breeding scheme was found to be an effective strategy for increasing progeny TTY over the 4x parents. However, the high degree of heterosis for TTY along with good GTA scores observed in certain cross combinations derived from these unadapted 2x species was a somewhat surprising result. Thus far, the importance of these two South American wild potato species, from the potato‐breeding standpoint, has been limited to the fact that they are natural reservoirs of major genes controlling resistance against important diseases and insects. However, the level of performance of some 4x‐2x families in comparative assays with elite cultivars suggests the unanticipated possibility of introgressing genetic factors from S. berthaultii and S. tarijense with positive effects on quantitative traits of horticultural importance along with these major resistance genes.     

The importance of wild potato species resistant to the potato cyst nematode,Globodera pallida,pathotypes P4A and P5A,in potato breeding. I. Resistance studies

Summary Seven wild diploid potato species, Series Tuberosa, representing 1023 clones were screened for resistance to the potato cyst nematode, Globodera pallida. Over 25% of the clones were resistant to pathotype P₄A and almost 30% were resistant to pathotype P₅A. The resistance in hybrid progenies of these and other resistant species with cultivated potatoes was evaluated, and over 2200 seedlings were screened. High frequencies of resistance (>50%) to P₄A were found in progenies with Solanum leptophyes, S. vernei, S. gourlayi and S. capsicibaccatum, whereas resistance to P₅A was found in these species as well as S. sparsipilum. The importance of nematode resistant wild species for potato breeding is discussed.     

Characterization of resistance to Globodera rostochiensis pathotype Ro1 in cultivated and wild potato species accessions from the Vavilov Institute of Plant Industry

Potato cyst nematodes (PCN) collected in six localities in the Leningrad region of North West Russia were identified as Globodera rostochiensis pathotype Ro1 and were used for subsequent resistance tests. Seventy‐nine accessions of cultivated and closely related wild potato species from the VIR collection in Russia were screened on resistance to G. rostochiensis pathotype Ro1 and on the presence of molecular markers for H1 and Gro1‐4 resistance genes. No associations were detected between the resistance level of diploid and tetraploid Andean and tetraploid Chilean potato landraces (indigenous cultivars) and their related wild species and their geographical distribution or presence of PCR‐based markers that are associated with the H1 and Gro1‐4 genes. At the same time, all susceptible genotypes lacked such markers. New sources of resistance were found and could be used in breeding.     

甘薯羽状斑驳病毒海南分离物全基因组序列克隆及分析

为了明确海南本地甘薯羽状斑驳病毒(sweet potato feathery mottle virus,SPFMV)种群的遗传进化关系及致病机理,本研究利用Small RNA测序技术对采自海南的甘薯样品进行鉴定,获得与SPFMV序列具有相似性的85个contigs,与NC_001841相似性较高。试验设计了5对共8条引物,分别为SPFMV-1F/1R、SPFMV-11F/1R、SPFMV-2F/2R、SPFMV-3F/3R和SPFMV-3F/33R PCR,扩增获得3987 bp、3710 bp、1996 bp、3369 bp和4730 bp目的片段。在此基础上再分段设计引物,进行基因全序列PCR扩增,序列拼接获得SPFMV-HN基因组全序列,其包含完整编码阅读框。本研究结果首次报道SPFMV海南分离物基因组全长,并且在P1区预测到另一个外框元件(pretty interesting sweet potato potyviral ORF,PISPO),聚合酶滑移机制可以产生带有额外核苷酸的转录变体,这些核苷酸可以翻译为P1N-PISPO和P3N-PIPO。本研究结果为进一步探究SPFMV致病分子机理和培育抗病甘薯品种奠定研究基础。     

马铃薯不同品种(系)和稻、薯轮作模式对根结线虫病的防治效果

根结线虫病是马铃薯连作减产的主要原因之一。为研究根结线虫病的农艺防治措施,本文开展了品种抗性评价、稻、薯轮作和土壤淹水下的线虫消减动态研究。抗性评价试验表明, 18个品种(系)中没有筛选到同时抗南方根结线虫和爪哇根结线虫的材料,品系1002-1和品种丽薯6号相对抗性较好,青薯9号和合作88相对敏感。马铃薯根结线虫发病土壤中可同时检测到卵、幼虫和成虫等不同形态。水旱轮作试验表明,经一茬水稻栽培可使土壤中根结线虫虫态完全消失,再种植马铃薯也不再发生根结线虫病,且马铃薯有效产量显著提高,青薯9号、合作88和丽薯6号的单株有效产量较对照分别提高50.5%、43.7%和26.0%,差异均极显著, 1002-1单株有效产量与对照差异不显著。由此可见,稻、薯轮作根结线虫防治效果明显。为进一步探索水旱轮作的防治机制,观察了土壤淹水进程中各种虫态消减动态。结果表明,在淹水过程中,根结线虫的卵、幼虫和成虫不同虫态此消彼长,要淹水60 d以上,土壤中才检测不到所有形态虫体。水早轮作中水稻生长期淹水时间足够长,满足防治线虫的淹水条件。因此结合抗病品种选用进行稻、薯轮作是防治马铃薯根结线虫病既绿色环保又可持续有...     

Resistance to potato late blight (Phytophthora infestans (Mont.) de Bary) in Solanum nigrum,S. villosum and their sexual hybrids with S. tuberosum and S. demissum

Summary Solanum nigrum and S. villosum, and their sexual hybrids with S. tuberosum and S. demissum respectively, were inoculated with a complex race of Phytophthora infestans. No visible reaction was seen on S. villosum and one genotype of S. nigrum. Another genotype of S. nigrum and the hybrids showed a hypersensitive response on most inoculated leaves. In one experiment, some sporulation was observed on detached leaves of a hybrid derived from S. nigrum. Microscopical examination of infections in S. nigrum and in a hybrid from S. nigrum, showed that penetration of epidermal cells and subsequent intercellular growth of the pathogen into the spongy mesophyll occurred, but without the formation of haustoria, and that invaded and neighbouring cells became necrotic. Callose appositions were found in epidermis and mesophyll cells of all inoculated genotypes, and also in epidermal cells of the unrelated nonhost species Brassica campestris.     

Field resistance to late blight (Phytophthora infestans (Mont.) De Bary) in potato (Solanum tuberosum L.) somaclones associated with instability and pleiotropic effects

Summary Adventitious regenerants (somaclones) of Bintje and their vegetative progeny were screened for field resistance to Phytophthora infestans as follows: the area under the disease progress curve was computed and correlated with resistance rating in Bintje and reference varieties. The resistance rating of the somaclones was determined from this relationship.Clones with stable improved field resistance in successive years' trials were detected, however, most of such clones were also maturation mutants. Variation in resistance rating in clone replicates and between years was detected in most clones.The possible basis of the field resistance and reasons for its instability are discussed.Abbreviations AUDPC area under the disease progress curve     

The production of yellow-seeded Brassica napus (AACC) through crossing interspecific hybrids of B. campestris (AA) and B. carinata (BBCC) with B. napus

To transfer the genes for yellow seed coat from both genomes A and C to B. napus (AACC), the hexaploid of Brassica (AABBCC) was synthesised from reciprocal interspecific crosses between yellow-seeded B.campestris (AA) and B.carinata (BBCC). The hexaploid with 27 pairs of chromosomes was red-seeded which showed that genic interaction existed in the trigenomic plants for the colour of the seed coat. Hundreds of hybrid seeds were obtained from crosses between the red-seeded hexaploid and partial yellow or brown-seeded varieties of B. napus as pollen donor. The majority of the hybrid plants (AABCC) were self fertile with brown seeds. It appeared that the chromosomes of the B genome were excluded during the meiosis of the pentaploid and a high proportion of the genetically balanced AC gametes could be produced. The fertility of the F2 population was increased and even reached normal levels for some plants. Seventy-three plants with the yellow-seeded character were isolated from 2590 open-pollinated F2 plants, most with increased fertility. After two successive self-pollinations, 18 lines produced yellow seeds and no brown seeds segregated from these populations. The morphology of the novel yellow-seeded plants was basically towards B. napus. Esterase isoenzyme electrophoresis showed that the plants contained some of the genetic background of B. campestris, B. carinata and B. napus. Cytological analysis has shown that at least some yellow-seeded lines have the B.napus AACC genome composition with 38 chromosomes and normal meiotic pairing. This revised version was published online in July 2006 with corrections to the Cover Date.