Anthocyanin pigmentation in a potato dihaploid and its chromosome-doubled derivatives

Summary Tubers with lilac pigmented eyes were produced by tetraploids derived from vegetative chromosome doubling of a Pentland Crown dihaploid. Close examination of tubers of the original dihaploid revealed very faint pigmentation in one eye on each of two tubers. Although there is some capacity of the dihaploid to produce tuber eye anthocyanin pigment therefore, doubling the chromosome number had apparently resulted in increased anthocyanin production.     

The male fertility of primary and further generation dihaploids ofSolanum tuberosum

Summary Male sterility in dihaploids ofSolanum tuberosum is often a barrier to their utilization in breeding and genetical studies. Although the male fertility of primary dihaploids (obtained directly from tetraploids) was generally low, a few were produced by the author which produced seed when used as pollen parents on female fertile dihaploids. The population of further generation dihaploids (from dihaploid intercrosses) produced from the showed a marked improvement in male fertility. Comparative data from measurements of different aspects of male fertility in dihaploids are presented. These showed that the weight of pollen per anther, the percentage of (iodine) stainable pollen and the number of seeds per berry were greater in further generation dihaploids than in primary dihaploids. It is suggested that interspecific crosses using dihaploids are not necessarily useful for increasing male fertility in diploid potatoes. The advantages of breeding at the diploid level withinS. tuberosum are discussed. Rare male-fertile dihaploids could be used to generate diploids with a high frequency of male fertility. These could then be used to cross with any other flowering dihaploids to combine characters at the diploid level, so exploiting the simpler genetic ratios associated with disomic inheritance.     

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.     

Transfer of PLRV resistance fromSolanum verrucosum Schlechdt to potato (S. tuberosum L.) by protoplast electrofusion

Summary Somatic fusions between an accession of the diploid wild speciesSolanum verrucosum and a dihaploid S.tuberosum genotype were produced in order to incorporate resistance to potato leafroll virus (PLRV). In total 15 somatic hybrids out of 16 regenerants were obtained. Identification of hybrids was based on additive RAPD patterns, general morphological characteristics, chromosome numbers and chloroplast counts in stomata guard cells. A field trial was performed with the hybrids, their two parents and the control cultivar Kennebec to assess field performance and phenotypic variability. Yield parameters varied considerably among somatic hybrids. Some of the hybrids gave significantly higher yields, tuber numbers and tuber weights than both parents. Pollen fertility of hybrids ranged from 19 to 77%. Twelve hybrids were found to be resistant to PLRV.     

用组培法培养马铃薯—倍体和双单倍体进行染色体加倍的研究

分别取来自普通栽培种(S.tuberosum)的两个一倍体品系和两个双单倍体品系的茎段和叶片,用两步法进行组织培养来加倍其体细胞染色体数目。在以MS+2.25mg/1 BAP+5mg/1 NAA 为第一步的培养基上,从一倍体和双单倍体的茎段和叶片中获得了许多再生植株,其中约60%的加倍成植株(2n=2x=24或2n=4x=48),此结果表明此法是把单单倍体(以下简称一倍体)和双单倍体转化为纯合同源四倍体的有效方法。     

Pedigree,agronomic and molecular divergence of parents in relation to progeny performance in potato

Summary Inbreeding depression is often observed in potatoes (Solanum tuberosum L.). The influence of four types of genetic divergence on the vigour and variability of the progenies was studied in two field experiments. The measures of genetic divergence were 1) the progenies' inbreeding coefficients (F); 2) The Mahalanobis distances between the parents obtained from their agronomic traits (A); 3) the parental GCA effects (G); 4) quantitative measurements of proteins and esterases separated by PAGE (M). These four measures of divergence were not significantly related. The inbreeding coefficient was the only parameter which had significant simple correlations with total yield, total tuber number, SCA effects for total yield or the standardized generalized variance. Canonical correlation analyses between the divergence parameters and vigour-related traits produced significant relationships in one experiment only. Progeny with high inbreeding coefficient values (0.06–0.12) gave below average yield. It is suggested that an inbreeding coefficient should not exceed 0.08. None of the methods of estimating genetic divergence appears to be a good predictor of either the mean or the variability of a progeny.     

Bacterial wilt resistance in the progenies of the fusion hybrids between haploid of potato andSolanum commersonii

Bacterial wilt of potatoes, which used to be a widespread disease in tropics and subtropics, has become a threat to potato production in temperate region. The diploid species Solanum commersonii has several desirable characteristics including cold tolerance and resistance to several diseases. Selected somatic hybrids between S. tuberosum dihaploid and bacterial-wilt-resistant S. commersonii clones were cross pollinated with S. tuberosum cultivars for further selection of bacterial wilt resistance. The chromosome numbers of the fusion parents were confirmed as 24, and the three fusion hybrids crossed were all tetraploids. The chromosome number of 11 backcross 1 progenies (BC1) was 48 and that of the other six was close to the tetraploid number. Backcross 2 progenies (BC2) were obtained from only three of the 44 BC1 clones crossed. The S. commersonii parent clone, LZ3.2, was the most resistant to bacterial wilt among wild species clones tested. The first sexual progenies segregated for resistance, with one clone highly susceptible and four clones highly resistant. Three highly resistant BC1 clones, CT02-4, CT08-4, and CT10b-4, were backcrossed to cultivars. Two cross combinations produced mostly susceptible BC2 progenies; however seven clones were resistant or highly resistant for both race 1 and race 3. The highly resistant three clones, CT204-3, CT206-9 and CT206-10, were selected for the further testing as cultivars or breeding materials.     

Haploid,diploid and tetraploid parthenogenesis in interspecific crosses betweenSolanum tuberosum interdihaploids andS. phureja

Summary Among 70017 seeds in progenies of crosses from 23 interdihaploids with aS. phureja clone, homozygous for the seed marker ‘embryo spot’, 2 parthenogenetic monohaploid, 18 dihaploid and 3 tetraploid plants were detected. It is postulated that they have arisen from reduced, unreduced and doubled unreduced unfertilised egg cells.     

马铃薯加工性状亲子代间表现型差异的研究

试验选用20个具有良好加工性状表现的马铃薯品种或品系,配制了16个杂交组合,在实生苗当代进行子代测验,以验证亲本的表型和杂种实生苗当代块茎表现的关系。通过对后代群体块茎的比重、长宽比、芽眼深浅等性状的表现,来评价亲本和组配方式对后代性状表现的影响。结果表明:根据亲本表现在比重、薯形,芽眼三个性状上不能够完全准确地预测其子代表现,需要通过其子代表现来进行评价。在组合的配制效应上,Vester×Fl1867、Fl1771×台湾红皮、Wauseon×Caribe、以及CIP380854.3×Fl1533的后代群体表现好,可以作为选育加工品种的组合在育种应用。     

Oryzalin Treatment of Potato Diploids Yields Tetraploid and Chimeric Plants from which Euploids could be Derived by Callus Induction

Shoot apices from six diploid potato genotypes for polyploidisation were treated with oryzalin and colchicine solutions at different concentrations and incubation times. In addition to chimeric plants, diploids, one octoploid and completely ploidy doubled plants (4x) were regenerated in these experiments. Tetraploid plants were obtained for three of the six genotypes treated with oryzalin, and no tetraploid plants were obtained after colchicine treatments of any of the genotypes. The best result of 43% tetraploid plants was achieved using 28.8 μM oryzalin and 24 h of incubation with the L37 genotype. Moreover, we determined that doubled plants should not be selected on the basis of a single measurement and require additional ploidy checks in successive clonal generations. Some L37 plants that were initially identified as tetraploids by flow cytometry produced diploid or mixoploid plants in the next clonal generation. Morphological characters of doubled L37 plants were compared with the original genotype. The PVY resistance present in a parental clone was maintained in all regenerants and significant differences among ploidy variants were found for some quantitative characters. Molecular AFLP analyses did not show any differences among genotypes. Since polyploidizing substances may produce a large proportion of chimeric plants, we applied a method to increase the number of tetraploid plants. Callus culture of chimeric leaf explants was performed on regeneration medium and tetraploid shoots were obtained in four of the six genotypes tested. The best result of 40% tetraploid plants was achieved with explants from the L37 genotype. This method was successful in producing additional tetraploid genotypes from mixoploid plants obtained after oryzalin treatments.     

马铃薯4x×2x杂种无性一代产量及产量性状的表现

产生２ｎ花粉的Ｎｅｏ－ｔｕｂｅｒｏｓｕｍ双单倍体×Ｓｏｌａｎｕｍｐｈｕｒｅｊａ的二倍体杂种以及其与Ｎｅｏ－ｔｕｂｅｒｏｓｕｍ双单倍体的回交后代与普通马铃薯（Ｓ．ｔｕｂｅｒｏｓｕｍｓｓｐ．ｔｕｂｅｒｏ ｓｕｍ品种东农３０３进行了杂交。４ｘ杂种在株高方面表现出了很强的杂种优势。商品薯产量与４ｘ亲本无显著差异，但两个杂种群体的最高值分别超４ｘ亲本的２６０ｇ／株和６０ｇ／株。总产量杂种优势明显，但杂种单株结薯数多，平均块茎重小，这些是限制４ｘ×２ｘ杂种利用的不利因素，需要进一步地改善。     

The production of dihaploids and tetraploids with combined quantitative resistance to potato cyst nematode (Globodera pallida) and foliage blight (Phytophthora infestans)

Summary Dihaploids were produced from tetraploids which had resistance to foliage late blight (P. infestans) and had a range of levels of resistance to white potato cyst nematode (PCN) (G. pallida). Twelve of 33 dihaploids tested had useful levels of both resistances, 11 were resistant to blight alone and seven to PCN alone. There were no significant correlations between the resistances of the parents and the mean resistance of their dihaploids, for either character. There was also no correlation between levels of resistance to blight and to PCN. There were significant differences in mean levels of resistance to blight and to PCN between the tetraploid progenies of two dihaploids which possessed combined resistance.     

马铃薯种间体细胞杂种植株的细胞学观察和过氧化物同工酶谱分析

对马铃薯双单倍体品系81－15（2n＝2x－24）和南美二倍体栽培种Solanumphureja（2n＝2x＝24）体细胞融合获得的15个林系、81－15和二倍体野生种S．chacoense（2n＝2x＝24）体细胞融合获得的10个株系进行了细胞学鉴定和过氧化物同工酶谱分析。结果表明,杂种植株除一个株系为非整倍体外（染色体数为37）,其余株系均为四倍体（2n＝2x＝48）,是两个亲本的染色体数之和。与二倍体的双亲相比,杂种植株的叶下表皮气孔保卫细胞内叶绿体数目较多,但单位面积气孔数目较少,明显地表现出了四倍体的特征。杂种植株的过氧化物同工酶谱是其双亲酶谱谱带的总和。     

Correlation between resistance to late blight in foliage and tubers in potato clones from parents of contrasting resistance

Summary The relationship between resistance to late blight in foliage and tuber was examined on 50 clones of each of five progenies from crosses where one parent was resistant in both foliage and tuber and the other susceptible. Foliage resistance was assessed in a field trial and tuber resistance in a laboratory test on glasshouse-grown tubers. The genetical and environmental components of variance and the phenotypic and genetical correlations between foliage and tuber blight scores were determined for each progeny. Foliage and tuber resistance were correlated, indicating either that both are determined by the same genes or by different linked genes. The environmental component of variation was greater for tuber blight than foliage blight. We suggest that the most effective way of selecting for resistance to both aspects of the disease in a breeding programme is to select those resistant in the foliage and then screen them for tuber resistance.     

Insert copy number,chromosome number,pollen stainability,and crossability ofAgrobacterium-transformed diploid potato

Two 2n-pollen producing cultivated diploid, 2n = 24, potato genotypes DM56.4 and US-W5337.3 were inoculated with binaryAgrobacterium vectors. The T-DNA contained a neomycin phosphotransferase II coding region fused to a nopaline synthase promoter providing gene expression in plants and resistance to kanamycin. A total of 28 clones regenerated as adventitious shoots from callus tissue, growing on medium with kanamycin, was analyzed for chromosome number, pollen stainability, and crossability. Several clones were tested and found to retain crossability with diploid and tetraploid clones. Nine out of 10 of the regenerants from DM56.4 had been doubled; seven of these were tetraploids with 2n = 48, one clone had 2n = 49, and another 2n = 46. Five of 14 regenerants of US-W5337.3 had been doubled, and three of these had fewer than 2n = 48. Pollen stainability revealed that the tetraploid regenerants from DM56.4 were male sterile, whereas tetraploids from US-W5337.3 had stainable pollen. All of the diploid regénérants produced 2n pollen. Crossability of several diploid and tetraploid regenerants was high. The number of T-DNA inserts revealed by Southern blot analysis was also determined. Six, 10, and 9 regenerants had 1, 2, and 3 inserts of T-DNA, respectively. There were single regenerants each with 5, 6, or 7 inserts. Selection for higher insert number among regenerants should be feasible based on the range of insert number found. Identical restriction patterns within three groups of clones indicated that the 28 clones analyzed emerged from only 21 cells derived from independent transformation events. The variability in chromosome number and pollen stainability suggests that variation in other traits would be obtainable.     

对马铃薯4x-2x,2x-4x及2x-2x杂种四倍体进行花药培养获得转育2n配子基因的双单倍体新个体

利用3个2n配子材料(2x)在马铃薯(S.tuberosum L.)中进行4x—2x,2x—4x和2x—2x的杂交,获得了4个四倍体杂种材料;然后对它们进行花药培养,共得到32个双单倍体植株。检查其中23个植株,有2株是具5％以上2n花粉粒的双单倍体,1株是重组了2n卵基因的双单倍体。由此证明花药培养的倍性操作技术是转育马铃薯2n配子性状给双单倍体的有效方法之一。     

Cytoplasmic substitution inSolanum. II. Tuber characteristics of reciprocal backcross progeny

Summary Recurrent backcrosses (BC) were used to introduce nuclear factors ofS. tuberosum L. ssp.tuberosum into the cytoplasm of Andean potatoesS. tuberosum ssp.andigena (Juz. and Buk.) Hawkes andS. phureja Juz. and Buk. Each generation BC was produced in reciprocal directions to test the effect of cytoplasmic substitution on yield. This report analyses tuber number, tuber weight, and tuber length of BC₂ and BC₃ substitution into ssp.andigena cytoplasm and BC₂ and BC₂ substitutions intoS. phureja cytoplasm. Direction of cross had no consistent significant effect on yield components of ssp.andigena substitution lines. InS. phureja reciprocal progenies direction of cross was responsible for significant differences of some reciprocal sets. When there were significant differences the higher yield usually occurred when ssp.tuberosum was the pistillate parent, but in some progenies when ssp.tuberosum was the pistillate parent the yield was significantly lower than in the reciprocals. It is suggested that differences between reciprocals resulted from chromosomally encoded gene action, resulting from maternal and/or paternal effects, rather than cytoplasmic factors. Authorized for publication as paper No. 7494 in the Journal Series of The Pennsylvania Agriculture Experiment Station.     

双单倍体马铃薯染色体加倍的方法

本试验研究了马铃薯双单倍体(双单倍体是由四倍体栽培种花药培养产生的)植株加倍的两种方法。第一种方法:在MS培养基内附加NAA 0.1mg/1,而后加入秋水仙素0.2mg/l,用马铃薯双单倍体植株切断接种。经过40天培养,获得了加倍的四倍体植株,加倍频率为63.6%。第二种方法:将马铃薯双单倍体植株的茎,接种在含有2.4-D 1mg/l,萘乙酸0.025mg/l,KT 2mg/l的MS培养基上,在产生愈伤组织后诱导出再生四倍体植株,加倍频率为22.3%。     

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.     

Assessing progenies of potato for resistance to skin spot (Polyscytalum pustulans) in the glasshouse

Summary Crosses were made between six parents differing in susceptibility to skin spot and glasshouse-grown tubers from true seedlings of the nine resulting progenies assessed for resistance to the disease. The assessment was evaluated by a two-year comparison with the behaviour of field-grown tubers of the progenies and their parents. The correlations between progenies in the glasshouse and field were r=0.911 and 0.753 and those between the mid-parental values and the progeny means were high (r>0.864), indicating that parental differences were heritable. It is concluded that correlations were sufficient for glasshouse-grown tubers to be used for assessing large numbers of progenies in genetical studies and for eliminating highly susceptible progenies at an early stage in a breeding programme. R.L. Wastie died 16.01.96.