Evaluation of dihaploid populations from potato varieties and breeding lines

Summary Dihaploid populations from 20 varieties and 11 breeding lines ofSolanum tuberosum differed greatly in the occurrence and frequency of mutant phenotypes, in tuberization, flowering, pollen stainability, 2n-pollen production and resistance toGlobodera rostochiensis pathotype Ro-1. One population contained up to 61.7% lethal mutants: 39.1% of the total of 5377 dihaploids obtained did not tuberize, and 32.4% of 825 vigorous dihaploids did not flower. Of 825 vigorous dihaploids, 26.9 and 3.9% respectively had a pollen stainability higher than 20 and 60%. Among 222 dihaploids with a pollen stainability higher than 20%. 23.0% produced some 2n-pollen and none of these had a stainability higher than 60%. Among the dihaploid populations investigated the proportion of resistant to susceptible dihaploids ranged from 8:26 to 62:8.     

2n gametes inSolanum tuberosum dihaploids

Pollen stainability over two years and microsporogenesis were investigated in seven dihaploids extracted fromSolanum tuberosum tetraploids (AVRDC-1287.19, Conchita, Merrimack and W231). Two dihaploids proved to be male-sterile and male gamete abortion is likely to be post-meiotic because a regular meiosis occurred. The other dihaploids proved to be partially male-fertile and 2n pollen producers, as well. Almost all clones showed predominantly the formation of parallel spindles. Correlations estimated between the observed abnormalities in spindle orientation and the meiotic products evidenced a high correlation between parallel/fused spindles and dyads. The dihaploids were also used as parents in crosses with tetraploids. One berry with 44 seeds was obtained from dihaploid AVI 24 used as pollen parent in 4x × 2x crosses. After 2x × 4x crosses, three dihaploids including the two male-sterile were found to be 2n egg producers. The formation of 2n gametes inS. tuberosum dihaploids is discussed in relation to a direct use of dihaploids in sexual polyploidization schemes to combine useful traits in a superior genotype at 4x level.     

马铃薯新型栽培种(Neo-tuberosum,Solanum tuberosum ssp.andigena)双单倍体产生2n配子突变体的研究(一)

采用新型栽培种(Neo-tuberosum,2n=48)为亲本,以IVP_(35)作授粉者,诱发孤雌生殖,获得了148份双单倍体无性系,从中筛选出能够产生FDR型2n配子无性系16份(纺锤体平行并融合类型11份,不联会突变型5份)。由于双单倍体孕性低或败育,以入选的16份双单倍体直接同S.tuberosum品系或品种杂交未能成功。同类型(A)入选的双单倍体无性系的系内或系间授粉,因配子体同型复等位基因作用产生的自交不亲和,也没能得到实生种子,而以产生2n配子的S.phureja品系的混合花粉授粉,在5个组合中获得25个浆果,共825粒杂交种子。     

Effect of functional female and male fertilities on crossability in diploid potato breeding

Summary Functional female fertility (FFF) and functional male fertility (FMF) were studied in diploid potato clones, includingS. tuberosum haploids, F₁ hybrids between haploids and otherSolanum species, and in progenies from subsequent hybridisations. The results of 402 crosses between 91 diploid potato clones revealed that 60% of all genotypes had low levels of FFF as well as low levels of FMF regardless of their origin and degree of hybridity. Failed crosses (without any fruits at all) and those with a low level of success (with less than 3 seeds per pollinated flower) constituted 69% of all studied crosses. Most of these had resulted from parents with low levels of FFF and FMF, and these two characteristics were expressed independently. Functional female fertility affected crossability behaviour no less than FMF, therefore both FFF and FMF should be controlled in diploid potato clones involved in breeding. Reliable FFF and FMF estimates can be based only on crossing results. The coefficient of multiple correlation between seeds per pollinated flower in crosses and FFF and FMF estimates of the parental clones was 0.80 (P<0.01). Hence the compilation and analysis of the results of diploid potato crosses allow a reliable assessment of FFF and FMF and consequently the prediction of crossability with a degree of sufficient accuracy.     

Male fertility and freezing tolerance of hybrids involvingSolanum tuberosum haploids and diploidSolanum species

Summary Male sterility of (haploid x species) hybrids often hampers the exploitation of wildSolanum species in breeding for cold tolerance. In a study aimed at finding ways round these problems, we found that one accession of the cold tolerant wild speciesS. sanctae-rosae gave progenies segregating for male fertility in crosses withtuberosum haploids. Moreover, a source of male fertility restorer gene(s) was hypothesized in the (haploid x species) hybrid UP88-C80 (tbr x chc). A relatively high freezing tolerance was found in the progenies coming fromtuberosum haploid xsanctae-rosae crosses and some hybrids showed a freezing killing temperature as low as −5.4°C. Although the most tolerant clones did not shed pollen, at least one clone (Sr6,tbr x sct) combined male fertility, 2n pollen production and good freezing tolerance (−3.9°C) and might be directly used in crosses with tetraploidS. tuberosum in order to introgress resistance to low temperature intuberosum form. Contribution no. 137 from the Research Centre for Vegetable Breeding-CNR. Research supported by National Research Council of Italy, special project RAISA, subproject no. 2, paper no. 2776.     

马铃薯新型栽培种(Neo-tuberosum)双单倍体杂种的研究

利用马铃薯新型栽培种(Neo—tuberosum,2n=4x=48)的双单倍体(2n=2x=24)与S.phureja (2n=2x=24)杂交,克服新型栽培种双单倍体雄性败育获得了成功。鉴定结果表明,杂种实生苗绝大多数保持了二倍性(2n=2x=24);实生苗可育株占总数78.36％。经减数分裂观察,入选的17个DP优良单株系主要是SDR型2n配子。为了选育出FDR型2n配子突变系,加强DP优系中的新型栽培种的遗传成分,即以D_(2012)—22(SY—m型)为母本,以DP优系为父本配制了回交组合,得到1000多粒种子,供进一步研究利用。     

Diploid Potato Hybrids as Sources of Resistance to Blackspot Bruising

Resistance to bruising is an important quality trait of potato depending on multiple genetic and environmental factors. Thirty one interspecific diploid hybrids and two tetraploid standards were evaluated for resistance to bruising in 2 years. Diploids originated from crosses between wild and primitive Solanum species and dihaploids of S. tuberosum. Two-year mean values of bruising resistance of tested genotypes indicated significant variability of the trait. Analysis of variance showed significant effects of genotype, year and interaction between them on the trait. Broad-sense heritability of bruising resistance was estimated as moderately high, Hb?=?0.73. Content of L-tyrosine, the main substrate of discoloration reaction, was not significantly correlated with brusing in tested diploid hybrids. The clones, that were resistant to blackspot bruise, also combined quality traits and resistances to many pathogens. All these traits could be transferred together into tetraploid level in 4x?×?2x crosses via male 2n gametes produced by diploid hybrids.     

Investigation of dihaploid potatoes in the netherlands

Summary Two hundred primary dihaploids (2n=2x=24) were obtained relatively easily from Dutch varieties and breeding stocks of the potato (2n=4x=48). Male fertility is rare in primary dihaploids but increases rapidly with further crossing. It is thus possible to inbreed and cross-breedSolanum tuberosum at the diploid level. Tetraploids were obtained by colchicine treatment. Tetraploids were also obtained by backerosses of dihaploids with tetraploids.

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Zusammenfassung Die Untersuchungen mit dihaploiden Kartoffeln wurden im Jahre 1961 begonnen. Wir verfügen derzeit über eine Kollektion von etwa 200 dihaploiden. Darunter befinden sich Klone mit Resistenz gegenPhytophthora, Kartoffelkrebs und verschiedene Viren. M?nnliche Fertilit?t ist nur bei einem Klon gefunden worden. Indem wir amerikanische m?nnliche fertile Haploiden verwendeten (U.S.-W.), konnten wir eine Anzahl-Kreuzungen zustande bringen. Viele dieser Nachkommenschaften (sekund?re Dihaploiden) bezitzen guten Pollen. Einzelne sekund?re Dihaploiden sind selbstfertil. Die Nachkommenschaft zeigt eine starke Inzuchtdepression (Abb. 1). Dihaploide Klone, homozygot für bestimmte Eigenschaften, sind bereits aufgebaut worden. Tabelle 1 zeigt die Chromosomenanalyse einiger Nachkommenschaften in Valenzkreuzungen. Die so hervorgegangenen Tetraploiden sind m?nlich und weiblich fertil. Mit Hilfe von Colchizin kann die Chromosomenanzahl dihaploider Pflanzen verdoppelt werden. Diese Tetraploiden bilden aber keine Blüten.

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Résumé Les recherches sur pommes de terre dihaplo?des ont débutées en 1961. Depuis quatre années, nous disposons d'une collection de 200 dihaplo?des approximativement. II existe parmi ces génotypes des clones résistants auPhytophthora infestans, à la galle verruqueuse et à différentes maladies à virus. La fertilité male n'a été constatée que chez un seul clone. On a pu réaliser un certain nombre de croisements en utilisant des haplo?des “males fertiles” américains (US-W). Beaucoup de descendances secondaires (dihaplo?des secondaires) possèdent du bon pollen. Rarement les dihaplo?des secondaires sont autofertiles. La descendance montre un effet marqué de consanguinité (Fig. 1). On a déjà réalisé des clones dihaplo?des homozygotes pour certains caractères. Tableau 1 mentionne les résultats de l'analyse des nombres chromosomiques de quelques descendances de croisements de génotypes de plo?dies différentes. Les tétraplo?des obtenus sont également “males et femelles fertiles”. On peut doubler le nombre de chromosomes des plantes dihaplo?des par action de la colchicine chimique. Cependant ces tétraplo?des ne produisent pas de fleurs.

     

Cytoplasmic substitution inSolanum. I. Seed production,germination, and sterilities of reciprocal backcross generations

Summary Genes ofSolanum tuberosum L. ssp.tuberosum were introduced into cytoplasm ofS. tuberosum ssp.andigena (Juz. & Buk.) Hawkes andS. phureja Juz. & Buk. to reduce cytoplasmic sterilities. Alleles that influenced berry set and seed content produced differences between reciprocal progenies. Their expressions were interpreted in terms of genes of the maternal and/or paternal parents. When the ssp.andigena genotype was introduced from the maternal parent if favoured high berry set and low seed number. Maternal ssp.tuberosum favoured lower berry set and higher seed number. The source ofS. phureja cytoplasm, aS. phureja×S. chacoense F₁, used as maternal parent favoured lower berry and seed set. During successive backcrosses progeny expressions approached those of the recurrent ssp.tuberosum pollen parents. A low level of pollen sterility occurred in BC₂ plants, unrelated to direction of cross or cytoplasmic factors. By BC₄, seed set and fertility were as good as ssp.tuberosum, and recurrent backcrossing could be terminated without loss of seed production or fertility. Pennsylvania Agricultural Experiment Station Journal Series No. 7493.     

敲除TMS5基因获得温敏不育粳稻新材料

【目的】水稻温敏不育系的选育是两系杂交稻育种工作中的关键环节。TMS5是控制温敏不育的重要基因,在生产上也应用较广。为了加快水稻温敏不育系的选育进程,【方法】利用CRISPR/Cas9技术,在武运粳7号背景下对TMS5基因进行编辑。【结果】通过测序从22株T0代中筛选获得6株纯合的突变体,分析发现6个纯合的突变体产生了相同的遗传变异,都在TMS5基因第2外显子44 bp和45 bp之间插入碱基"A",导致翻译提前终止。通过细胞学观察和人工辅助授粉证实获得的株系花粉败育而雌配子正常发育。【结论】这些结果表明对温敏不育基因TMS5基因进行编辑可以快速获得粳型温敏雄性不育系。     

敲除TMS5基因获得温敏不育粳稻新材料

【目的】水稻温敏不育系的选育是两系杂交稻育种工作中的关键环节。TMS5是控制温敏不育的重要基因，在生产上也应用较广。为了加快水稻温敏不育系的选育进程，【方法】利用CRISPR/Cas9技术，在武运粳7号背景下对TMS5基因进行编辑。【结果】通过测序从22株T0代中筛选获得6株纯合的突变体，分析发现6个纯合的突变体产生了相同的遗传变异，都在TMS5基因第2外显子44 bp和45 bp之间插入碱基“A”，导致翻译提前终止。通过细胞学观察和人工辅助授粉证实获得的株系花粉败育而雌配子正常发育。【结论】这些结果表明对温敏不育基因TMS5基因进行编辑可以快速获得粳型温敏雄性不育系。     

二倍体马铃薯主要农艺性状及其2n花粉频率的表现

通过田间种植,比较了马铃薯二倍体与四倍体的长势、叶色、薯形、薯皮色、薯肉色、芽眼深浅、单株产量、单株块茎数、平均块茎重等植物学和农艺性状,并对二倍体马铃薯的花粉育性以及２ｎ花粉的频率进行了鉴定。根据单株产量、块茎数、薯形、２ｎ花粉频率等综合性状,筛选出ＤＹ１４－４－２５、ＤＹ４－３０以及Ｑ４９－１－９,为进一步开展马铃薯分解育种工作打下物质基础。     

Aneuploid hybrids from 5x − 4x crosses in potato: Chromosome number, fertility, morphology and yield

The objective of this study was to characterize sixS. commersonii - S. tuberosum progenies deriving from 5x ? 4x crosses to provide evidence that they can be used in potato breeding. Hybrids analyzed (coded PTH) had a chromosome number between the 4x and the 5x level. In particular, progeny means for chromosome number ranged from 50±0.5 to 54±0.7, with most (70%) genotypes exhibiting a low aneuploid level of 48 to 53 chromosomes. Despite being aneuploid, the hybrids did not generally show phenotypic aberrations or vigor reduction common to aneuploids of other species. Most genotypes resembledS. tuberosum in growth habit, whereas corolla type, eye depth, and stolon length varied between and within progenies. Aneuploid hybrids produced tubers under long-day conditions. However, tuber yields were not as high as expected, probably due to lack of adaptation of theS. commersonii genome to long-day conditions. Although aneuploidy has often been associated with reduced male and female fertility, many 5x ? 4x hybrids were fertile in crosses withS. tuberosum. The average berry set and number of seeds/berry were 33% and 34.7, respectively, following PTH ×S. tuberosum crosses, and 40% and 51.7, respectively, forS. tuberosum × PTH crosses. The useful variation for fertility and tuber traits found in this material is being used for further breeding efforts.     

马铃薯极早熟品种东农303 2n卵孤雌生殖无性繁殖系变异的研究

利用二倍体栽培种(S. phureja)“授粉者”(ⅣP35)诱导马铃著极早熟品种东农303 2n卵孤雌生殖获得四倍体无性繁殖系20余份。根据著肉、薯肉色的分离比例确定为第2次分裂重组核(SDR)的2n卵。此外,块茎芽眼的深浅、花粉育性和匍匐枝的长度等也有明显的性状分离。无性系PT_(3-3)、PT_(3-5)、PT_(3-6)和PT_(4-5)的花粉育性良好并产生自交实生种子以及不同无性系间的杂种实生种子。无性系PT_(3-3)和PT_(3-5)等具有一些优于亲本东农303的经济性状,如芽眼浅,薯形圆整,单株块茎产量较高。因此,由雄性不育的早期品种2n卵(SDR)孤雌生殖产生的四倍体无性系可作为选育早熟新品种的育种材料。     

小麦温敏不育系BNS366育性研究

为了揭示小麦温敏不育系BNS366的育性转换规律,为不育系的转育及杂种优势利用提供依据,于2011-2013年度连续2年采用大田分期播种试验分析了BNS366、BNS、矮抗58和扬麦13的育性特点。结果表明,随着播种期的推迟,BNS366表现出规律性育性转换特征。9月23日至10月17日播种,花粉败育率为99.77%~100%,自交不育度为99.21%~99.81%,每穗自交实粒数为0.10~0.51。10月25日播种,花粉败育率为91.78%~98.89%,自交不育度为92.86%~94.96%,每穗自交结实粒数为3.20~4.37,12月4日播种可育。BNS366具有两个育性转换的临界期,10月25日播期为不育变为高度不育的育性转换临界期,12月4日播期为半不育转换为可育的育性转换临界期,不育期花粉以典型败育为主。BNS366可以作为小麦杂种优势利用新的不育系。     

Differences between hybrids ofSolanum tuberosum L. andSolanum circaeifolium Bitt. obtained from symmetric and asymmetric fusion experiments

Summary Potato plants obtained by symmetric and asymmetric protoplast fusion experiments withSolanum tuberosum andSolanum circaeifolium were compared and their potential use for plant breeding was investigated. For asymmetric hybridisation wild species tissue was irradiated with X-rays. Irradiation enhanced regeneration frequency. RFLP analysis showed that both techniques gave approx. 95% hybrids among the regenerates. Asymmetric protoplast fusion gave 22% regenerates with a ploidy level between 2x and 4x or 4x and 6x. None of the regenerates obtained from symmetric fusion had these ploidy levels. The tetraploid hybrids showed good male and female fertility. Male fertility was highest for the eutetraploid regenerates. With increasing asymmetry and with increasing ploidy level fertility decreased. Berry induction and seed production were observed for the eutetraploid and, to a lesser degree, for the hypotetraploid and hypohexaploid hybrids.     

Resistance toErwinia spp. in diploid potato with a high starch content

Summary Tubers of diploid interspecific hybrids, hexaploid somatic hybrids ofSolanum tuberosum andSolanum brevidens, a fusion parental line (S. tuberosum) and several Polish and American commercial cultivars were screened for resistance toErwinia carotovora subsp.atroseptica andErwinia chrysanthemi, in laboratory tests. Both groups of hybrids showed a high level of resistance to soft rot and gave a similar type of resistant reaction, which distinguished them from the cultivars and fusion parent. Tubers of the diploid interspecific hybrids had a high starch content. Our results indicate the possibility of obtaining breeding material with a high starch content and resistance to bacterial soft rot.     

Male sterility and 2n pollen in 4x progenies derived from 4x×2x and 4x×4x crosses in potatoes

Summary The objectives of these experiments were to evaluate male sterility (MS) and 2n pollen frequency in 77 4x×2x families and 26 4x×4x families. The 2x parents were haploid-species hybrids and the 4x parents mainly clones withS. tuberosum ssp.tuberosum (tbr) andS. tuberosum ssp.andigena (adg) in their genetic background. The female parents had different cytoplasms: adg, tbr,S. demissum (dms), andS. stoloniferum (sto). Families from female parents with adg or dms cytoplasm did not have MS plants. However, families derived from crosses of many tbr females with 2x hybrids and a 4x adg clone had a high percentage of MS plants; all the progenies from cv. Serrana (sto cytoplasm) possessed tetrad sterility. These results can be explained as due to an interaction between a dominant male sterility (Ms) gene with tbr and sto cytoplasms. Some MS 4x hybrids (tbr×adg) used as females had the Ms gene, but male fertility was restored in some plants of their progenies when they were crossed with 4x tbr clones. This indicates that some tbr clones have a fertility restorer gene (Rt). The results from both 4x×2x and 4x×4x families fit the expected ratios assuming chromatid segregation for both the Ms and Rt loci. The gene frequency of parallel spindles was estimated in the 4x population as 0.74.     

Fertility of doubled monoploids ofSolanum phureja

Thirteen anther-derived doubled monoploids and their heterozygous diploid anther donor were evaluated in the greenhouse for flowering, fruit set, seed set and pollen fertility. Doubled monoploids varied for number of days to flower, duration of flowering, abundance of flowers and general flower quality. Fruit set on the doubled monoploids in controlled crosses with diploid male-fertile clones varied from 0 to 73% with a mean of 33% compared to the anther donor with 58%. Seed set varied from 9 to 210 seeds per fruit and was significantly lower than the anther donor (377 seeds/fruit). Five doubled monoploids that shed low quantities of pollen exhibited significantly less pollen stainability (10-30%) than the anther donor (88%). However, pollen of doubled monoploids did not germinatein vitro. 2n pollen frequencies of 20–55% were observed for the doubled monoploids compared with 22% for the anther donor. It was concluded that doubled monoploids had lower female fertility compared to the anther donor but that they produced sufficient seed to facilitate utilization in a breeding program as female parents. However, they were considered to be male-sterile for practical purposes.     

Triploids of the common potato,Solanum tuberosum L.

Summary Triploid plants of the common potato (Solanum tuberosum L. group Tuberosum) were derived from crosses between tetraploid varieties and selected dihaploids. The ability to induce triploidy seems to be largely dependent on the genotype of the dihaploid parent. Morphologically there was very little difference between triploids and their tetraploid siblings. Yields of triploids could be equivalent to genetically similar tetraploids.