Fertilization fitness and offspring ploidy in 3x × 2x matings in potato

Abstract

The main objective of the current research was to study the reproductive behaviour of artificial triploid potato hybrids between wild Solanum commersonii and the cultivated potato Solanum tuberosum. When used in 3x × 2x crosses, triploids gave aneuploid progenies with somatic chromosome number ranging from 29 to 36. Fertilization fitness data suggested that the survival rate of gametes produced by the triploid parents may be related to their chromosome number. In addition, consistent with molecular data, our results indicated that fitness of gametes and chromosome number of progenies are influenced by the genome dosage of interspecific triploids. Since a main route to polyploidy formation is via 2n gametes and triploids, our study may contribute to a better understanding of polyploid plant reproduction, evolution and breeding.     

Evolution of nematode-resistant <Emphasis Type="Italic">Mi</Emphasis>-<Emphasis Type="Italic">1</Emphasis> gene homologs in three species of <Emphasis Type="Italic">Solanum</Emphasis>

Plants have evolved several defense mechanisms, including resistance genes. Resistance to the root-knot nematode Meloidogyne incognita has been found in wild plant species. The molecular basis for this resistance has been best studied in the wild tomato Solanum peruvianum and it is based on a single dominant gene, Mi-1.2, which is found in a cluster of seven genes. This nematode attacks fiercely several crops, including potatoes. The genomic arrangement, number of copies, function and evolution of Mi-1 homologs in potatoes remain unknown. In this study, we analyzed partial genome sequences of the cultivated potato species S. tuberosum and S. phureja and identified 59 Mi-1 homologs. Mi-1 homologs in S. tuberosum seem to be arranged in clusters and located on chromosome 6 of the potato genome. Previous studies have suggested that Mi-1 genes in tomato evolved rapidly by frequent sequence exchanges among gene copies within the same cluster, losing orthologous relationships. In contrast, Mi-1 homologs from cultivated potato species (S. tuberosum and S. phureja) seem to have evolved by a birth-and-death process, in which genes evolve mostly by mutations and interallelic recombinations in addition to sequence exchanges.     

Ploidy manipulation of the gametophyte, endosperm and sporophyte in nature and for crop improvement: a tribute to Professor Stanley J. Peloquin (1921�C2008)

Background

Emeritus Campbell-Bascom Professor Stanley J. Peloquin was an internationally renowned plant geneticist and breeder who made exceptional contributions to the quantity, quality and sustainable supply of food for the world from his innovative and extensive scientific contributions. For five decades, Dr Peloquin merged basic research in plant reproduction, cytology, cytogenetics, genetics, potato (Solanum tuberosum) improvement and education at the University of Wisconsin-Madison. Successive advances across these five decades redefined scientific comprehension of reproductive variation, its genetic control, genetic effects, evolutionary impact and utility for breeding. In concert with the International Potato Center (CIP), he and others translated the advances into application, resulting in large benefits on food production worldwide, exemplifying the importance of integrated innovative university research and graduate education to meet domestic and international needs.

Scope

Dr Peloquin is known to plant breeders, geneticists, international agricultural economists and potato researchers for his enthusiastic and incisive contributions to genetic enhancement of potato using haploids, 2n gametes and wild Solanum species; for his pioneering work on potato cultivation through true seed; and as mentor of a new generation of plant breeders worldwide. The genetic enhancement of potato, the fourth most important food crop worldwide, benefited significantly from expanded germplasm utilization and advanced reproductive genetic knowledge, which he and co-workers, including many former students, systematically transformed into applied breeding methods. His research on plant sexual reproduction included subjects such as haploidization and polyploidization, self- and cross-incompatibility, cytoplasmic male sterility and restorer genes, gametophytic/sporophytic heterozygosity and male fertility, as well as endosperm dosages and seed development. By defining methods of half-tetrad analysis and new cytological techniques, he elucidated modes, mechanisms and genetic controls and effects of 2n gametes in Solanum. Ramifications extend to many other crops and plants, in both basic and applied sciences.

Achievements

Based upon a foundation of genetics, cytogenetics and plant reproductive biology, Dr Peloquin and co-workers developed methods to use 2n gametes and haploids for breeding, and used them to move genes for important horticultural traits from wild tuber-bearing Solanum species to cultivated potato for the betterment of agriculture. The resulting potato germplasm included combinations of yield, adaptation, quality and disease resistance traits that were previously unavailable. This elite plant germplasm was utilized and distributed to 85 countries by the CIP, because it not only increased potato yields and quality, it also broadened the adaptation of potato to lowland tropical regions, where humanity has benefited from this addition to their food supply.Key words: 2n gametes, endosperm balance number, haploid, Solanum, true potato seed     

植物2n配子发生及其遗传标记研究进展

文章综述了植物2n配子发生及其遗传标记研究现状,论述涉及有关形态学标记、细胞学标记、同工酶标记乃至DNA标记等遗传标记在2n配子研究中的应用,指出通过花粉形态观察、大小孢子母细胞减数分裂行为观察、杂种后代倍性鉴定以及亲子分子标记相关分析等,对2n配子发生、2n配子遗传类型与杂合性以及2n配子在育种实践中的有效性等进行研究。分子标记技术已经成为2n配子形成相关基因分析等方面的有力工具,利用分子标记等手段,研究2n配子形成的分子机理以及应用于辅助育种等将成为今后的热点,进而推动植物育种尤其是多倍体育种蓬勃发展。     

Ploidy level manipulations in potato through sexual hybridisation

There is no better use of sexual reproduction in regard to breeding and genetic research than the ploidy level manipulations possible in the potato and its relatives. Unique reproductive characteristics of tuber‐bearing Solanum species make possible: the production of gametes with unreduced chromosome number; the presence of an endosperm dosage system that regulates success of interploidy/interspecific crosses; the possibility to easily extract maternal haploids following crosses with S. phureja. This paper reviews results obtained in scaling genomic multiples up and down in potato, and relates these manipulations to breeding strategies for the genetic improvement of the cultivated potato. Several ploidy series have been developed, ranging from the monoploid to the hexaploid level. Cultivated tetraploids were scaled down to the diploid and monoploid level by haploidy. Scaling upward was achieved by sexual polyploidisation via 2n gametes that resulted in triploid, tetraploid, pentaploid, and hexaploid genotypes with a broad genetic base. Altogether, the success of ploidy level manipulations constitutes further proof that sexual polyploidisation played an important role in the polyploid evolution of Solanum species, and supports the idea that gene flow can be relatively easily accomplished through interploid and bridge crosses.     

Using disomic 4x(2EBN) potato species' germplasm via bridge species Solanum commersonii.

The cultivated potato Solanum tuberosum Dunal has many wild related species with desirable traits. Some of these wild tetraploids have disomic chromosome pairing, ready selfing with little inbreeding depression, but have strong crossing barriers with cultivars. They hybridize most easily with 2EBN forms (which include most diploid species). Chromosome doubling to the 8x level, use of 2n gametes, use of 2n gametes of 4x-2x triploid hybrids, and embryo rescue have been proposed to overcome the crossability barrier of these species with S. tuberosum. In this study, 2x S. commersonii (cmm) was used as a bridge species with S. acaule and series Longipedicellata species. Synthetic tetraploid 4x-cmm crossed readily to disomic 4x species, resulting in fertile F1 and F2 hybrids. Some of these had 2n gametes, which enabled direct crossing to tuberosum, resulting in 6x hybrids. The benefits of this scheme are (i) hybrids are relatively fertile, so many progeny may be produced for selection at each step, (ii) hybridization with cmm results in 2n gametes needed for crossing to tuberosum, and breaks up restricted recombination within disomic genomes, and (iii) simple techniques and tools are employed.     

Cytogenetic analysis of Lycopersicon esculentum (+) Solanum etuberosum somatic hybrids and their androgenetic regenerants

The aim of the study was to characterize genomic relationships among cultivated tomato (Lycopersicon esculentum Mill.) (2n=2x=24) and diploid (2n=2x=24) non-tuberous wild Solanum species (S. etuberosum Lindl.). Using genomic in situ hybridization (GISH) of mitotic and meiotic chromosomes, we analyzed intergeneric somatic hybrids between tomato and S. etuberosum. Of the five somatic hybrids, two plants were amphidiploids (2n=4x=48) mostly forming intragenomic bivalents in their microsporocytes, with a very low frequency of multivalents involving the chromosomes of tomato and S. etuberosum (less than 0.2 per meiocyte). Tomato chromosomes showed preferential elimination during subsequent meiotic divisions of the amphidiploids. Transmission of the parental chromosomes into microspores was also evaluated by GISH analysis of androgenic plants produced by direct embryogenesis from the amphidiploid somatic hybrids. Of the four androgenic regenerants, three were diploids (2n=2x=24 or 2n=2x+1=25) derived from reduced male gametes of the somatic hybrids, and one plant was a hypertetraploid (2n=4x+4=52). GISH revealed that each anther-derived plant had a unique chromosome composition. The prospects for introgression of desirable traits from S. etuberosum into the gene pool of cultivated tomato are discussed. Received: 2 August 2000 / Accepted: 4 December 2000     

The complete chloroplast genome sequences of Solanum tuberosum and comparative analysis with Solanaceae species identified the presence of a 241-bp deletion in cultivated potato chloroplast DNA sequence

The complete nucleotide sequence of the chloroplast genome of potato Solanum tuberosum L. cv. Desiree was determined. The circular double-stranded DNA, which consists of 155,312 bp, contains a pair of inverted repeat regions (IRa, IRb) of 25,595 bp each. The inverted repeat regions are separated by small and large single copy regions of 18,373 and 85,749 bp, respectively. The genome contains 79 proteins, 30 tRNAs, 4 rRNAs, and unidentified genes. A comparison of chloroplast genomes of seven Solanaceae species revealed that the gene content and their relative positions of S. tuberosum are similar to the other six Solanaceae species. However, undefined open reading frames (ORFs) in LSC region were highly diverged in Solanaceae species except N. sylvestris. Detailed comparison was identified by numerous indels in the intergenic regions that were mostly located in the LSC region. Among them, a single large 241-bp deletion, was not associated with direct repeats and found in only S. tuberosum, clearly discriminates a cultivated potato from wild potato species Solanum bulbocastanum. The extent of sequence divergence may provide the basis for evaluating genetic diversity within the Solanaceae species, and will be useful to examine the evolutionary processes in potato landraces.     

The occurrence and frequency of 2n pollen in three diploid solanums from Northwest Argentina

Summary A group of wild, tuber-bearing species from Northwest Argentina, belonging to the series Tuberosa, Solarium spegazzini Bitt. (spg, 2n=2x=24), S. gourlayi Hawkes (grl, 2n=2x=24 and 2n=4x=48) and S. oplocense Hawkes (opl, 2n=6x=72), and Cuneolata, S. infundibuliforme Phil (ifd, 2n=2x=24), is being used to investigate the mode of origin of polyploids in the genus Solanum. 2n gametes have been detected in the diploid species ifd and spg and in a diploid race of grl, using cytological and breeding approaches. Twenty-two introductions of spg, 8 of grl and 26 of ifd have been tested for 2n pollen; 59%, 63% and 54% of them, respectively, had at least one 2n pollen producing plant. These introductions comprised 238, 76 and 235 plant respectively, of which 20, 16, and 32 plant produced 5% or more 2n pollen. The mechanism of 2n pollen formation was determined in several plant of 2x spg, 2x grl and 2x ifd. All of them were found to form diplandroids via parallel spindles. This mechanism, which gives meiotic products genetically equivalent to first division restitution gametes, is under control of the Mendelian recessive ps. The results suggest that the allele ps is widely distributed in natural populations of the three diploids, and that its frequency is very high. These species are seen as valuable material for population genetic studies, and for the eventual incorporation into a breeding scheme involving sexual polyploidization via 2n gametes.     

Effects of 2n-pollen formation by first meiotic division restitution with and without crossover on eight quantitative traits in 4x-2x potato progenies

 The 2n-pollen grains formed by first-division restitution without crossover (FDR-NCO) are unique breeding tools, since they can transmit almost 100% of non-additive genetic effects from the parent to the progeny. FDR-NCO gametes are considered superior to those formed by FDR with crossing over (FDR-CO), which can pass on to the progenies approximately 80% of the heterozygosity and a large fraction of the epistasis. However, 2n-pollen formation by FDR-NCO mechanism requires the incorporation (in homozygous condition) of at least two recessive alleles. In the present work, 40 tetraploid families derived from complete 4x-2x factorial crosses were evaluated under short-day conditions to verify whether or not the postulated genetic superiority of FDR-NCO over FDR-CO gametes holds true for eight quantitative traits in potato. Families were derived from crosses between four 4x commercial cultivars, and a random sample of ten diploid Solanum phureja-haploid S. tuberosum hybrids producing 2n-pollen by either FDR-CO or FDR-NCO. The results indicated no significant superiority of FDR-NCO over FDR-CO families for total tuber yield (TTY) and six other traits (haulm maturity – HM; plant vigor – PV; plant uniformity – PU; eye depth – ED; number of tubers per hill – NTH; and commercial over total yield index – CTI). Based upon cytological observations, the FDR-CO and FDR-NCO gametes are expected to be genetically equivalent for all loci between the centromeres and the chromosomal site of maximum recombination. In our experiment, differences between FDR-CO- and FDR-NCO-derived progenies were not observed for TTY. Therefore, our results can be interpreted as additional evidence for the hypothesis that genes with major effect on TTY expression might have a physical location between centromeres and proximal crossovers in the potato chromosomes. In addition, a similar trend was observed for HM, PV, and ED but apparently not for commercial yield – CY (i.e., tubers with more than 33 mm in diameter). Received: 21 September 1998 / Accepted: 26 October 1998     

Modulation of gene expression in cold-induced sweetening resistant potato species <Emphasis Type="Italic">Solanum berthaultii</Emphasis> exposed to low temperature

Cold-induced sweetening (CIS) is a crucial factor influencing the processing quality of potato tubers. To better understand the molecular events of potato CIS and different CIS-sensitivity among various potato species, a suppression subtractive hybridization library and cDNA microarray gene filters were developed. A total of 188 genes were found to be differentially expressed (DE) in Solanum berthaultii (ber) upon cold stimulation. These functional genes were mostly related to cell rescue, defense and virulence, metabolism, energy and protein fate, included in various processes of plant defense against abiotic stresses. Four expression patterns of these DE genes were profiled by qRT-PCR using the cold-stored tubers of both CIS-resistant (ber) and CIS-sensitive (E-potato 3, a variety of S. tuberosum) potatoes. The expression pattern and abundance of many DE genes encoding proteins involved in metabolism were different in these two potato tubers, especially genes associated with amylolysis, sucrose decomposition and glycolysis pathways, indicating distinct regulatory mechanisms between ber and E3 in response to cold stress, which may be crucial for potato CIS. Further investigation of these cold-regulated genes will deepen our understanding of the regulatory mechanisms of potato CIS and direct approaches for the genetic improvement of potato processing quality.     

Origin of chloroplast DNA diversity in the Andean potatoes

Summary Wide chloroplast DNA (ctDNA) diversity has been reported in the Andean cultivated tetraploid potato, Solanum tuberosum ssp. andigena. Andean diploid potatoes were analyzed in this study to elucidate the origin of the diverse ctDNA variation of the cultivated tetraploids. The ctDNA types of 58 cultivated diploid potatoes (S. stenotomum, S. goniocalyx and S. phureja), 35 accessions of S. sparsipilum, a diploid weed species, and 40 accessions of the wild or weed species, S. chacoense, were determined based on ctDNA restriction fragment patterns of BamHI, HindIII and PvuII. Several different ctDNA types were found in the cultivated potatoes as well as in weed and wild potato species; thus, intraspecific ctDNA variation may be common in both wild and cultivated potato species and perhaps in the higher plant kingdom as a whole. The ctDNA variation range of cultivated diploid potatoes was similar to that of the tetraploid potatoes, suggesting that the ctDNA diversity of the tetraploid potato could have been introduced from cultivated diploid potatoes. This provided further evidence that the Andean cultivated tetraploid potato, ssp. andigena, could have arisen many times from the cultivated diploid populations. The diverse but conserved ctDNA variation noted in the Andean potatoes may have occurred in the early stage of species differentiation of South American tuber-bearing Solanums.     

Genetical dissection of H3-mediated polygenic PCN resistance in a heterozygous autotetraploid potato population

Potato Cyst Nematodes (PCN) currently represent a serious threat to potato cultivation. However, many sources of resistance are known amongst primitive and wild relatives of cultivated potato, Solanum tuberosum ssp. tuberosum. Currently, in the UK, the major threat is due to Globodera pallida, resistance to which has not yet been effectively deployed in potato cultivars. We have performed linkage and QTL analysis of a tetraploid potato population segregating for the H3 source of resistance to G. pallida that was introgressed into cultivated potato from the primitive species, Solanum tuberosum ssp. andigena. This source is highly effective against the most common UK pathotype of G. pallida (Pa2/3) and its deployment in breeding material is a major goal. We adopted an approach involving bulked segregant analysis (BSA) as well as genome wide linkage analysis using AFLP and SSR markers. BSA provided a concentration of markers linked in coupling to a QTL on linkage group IV, and improved the accuracy of the QTL localisation. By performing an analysis on residual scores after removal of the effects due to the major QTL, we detected a second QTL on linkage group XI.     

Chromosome painting in meiosis reveals pairing of specific chromosomes in polyploid <Emphasis Type="Italic">Solanum</Emphasis> species

Analysis of chromosome pairing has been an important tool to assess the genetic similarity of homologous and homoeologous chromosomes in polyploids. However, it is technically challenging to monitor the pairing of specific chromosomes in polyploid species, especially for plant species with a large number of small chromosomes. We developed oligonucleotide-based painting probes for four different potato chromosomes. We demonstrate that these probes are robust enough to monitor a single chromosome throughout the prophase I of meiosis in polyploid Solanum species. Cultivated potato (Solanum tuberosum, 2n?=?4x?=?48) is an autotetraploid. We demonstrate that the four copies of each potato chromosome pair as a quadrivalent in 66–78% of the meiotic cells at the pachytene stage. Solanum demissum (2n?=?6x?=?72) is a hexaploid and has been controversial regarding its nature as an autopolyploid or allopolyploid. Interestingly, no hexavalent pairing was observed in meiosis. Instead, we observed three independent bivalents in 83–98% of the meiotic cells at late diakinesis and early metaphase I for the four chromosomes. These results suggest that S. demissum has evolved into a cytologically stable state with predominantly bivalent pairing in meiosis.     

Strategy differences of two potato species in response to nitrogen starvation. Do plants have a genetic switch for nitrogen signalling?

Survival responses to nitrogen starvation are well known in micro‐organisms but little studied in plants. To construct a framework for study of the plant responses, we investigated the strategy differences of tubers from two closely related potato species. Solanum tuberosum conserves tuber nitrogen by inhibiting shoot growth, but S. phureja mobilizes tuber nitrogen to grow shoots, flowers and seeds. Genetic analysis of progeny from S. phureja–haploid S. tuberosum crosses uncovered segregation of a single dominant gene for the S. tuberosum inhibition strategy. Within S. tuberosum, haploid progeny closely resembled their tetraploid parents, suggesting strong genetic control of the inhibition. Growth of the inhibited shoots was proportional to sub‐optimal levels of added nitrate, and was triggered by exogenous gibberellic acid (GA₃). These observations support the notion that potato plants can closely tie shoot growth to ambient nitrogen levels – probably by a root–shoot nitrogen signal transduction pathway, and that this can be overridden by emergency mobilization of nitrogen reserves, perhaps by GA signalling from the tuber. Furthermore, genes for such developmental switches can be identified by classical genetic analysis of closely related species, such as S. tuberosum and S. phureja, that exhibit opposite survival strategies.     

Identification of RFLP markers closely linked to the H1 gene conferring resistance to Globodera rostochiensis in potato

Summary Resistance to the root cyst nematode Globodera rostochiensis is an agronomic trait that is at present incorporated into most new potato varieties. Major dominant genes are available that originate from wild and cultivated Solanum species closely related to the cultivated European potato (Solanum tuberosum ssp. tuberosum). One of those genes, H1, from S. Tuberosum ssp. andigena, was mapped to a distal position on potato chromosome V using restriction fragment length polymorphism (RFLP) markers. The H1 locus segregates independently from Gro1, a second dominant gene presumably from S. Spegazzinii that confers resistance to G. Rostochiensis and which has been mapped to chromosome VII. One marker, CP113, was linked without recombination to the H1 locus.     

Reproductive ecology and genetic variability in natural populations of the wild potato,Solanum kurtzianum

The cultivated potato (Solanum tuberosum ssp. tuberosum) has more than 200 related wild species distributed along the Andes, adapted to a wide range of geographical and ecological areas. Since the last century, several collection expeditions were carried out to incorporate genetic variability into the potato germplasm around the world. However, little is known about the reproductive ecology and genetic population structure of natural potato population from field studies. The aim of this work is to study, in the field, the genetic variability and reproductive strategies of populations of one of the most widely distributed potato species in Argentina, Solanum kurtzianum, growing in Mendoza province. AFLP markers showed that the genetic variability is mainly present among plants within populations, indicating that in the sampled populations, sexual reproduction is more relevant than clonal multiplication (by tubers). Additional evidence was obtained evaluating the genetic diversity in populations with a distribution in patches, where several genotypes were always detected. From a field study performed in the Villavicencio Natural Reserve, we found that the average number of plump seeds per fruit was 94.3, identified and calculated the foraging distance of four insect pollinators, and demonstrated the seed dispersal by storm water channels. We argue that the breeding system, the two modes of reproduction and the ecological interaction described here may have a prominent role in determining the genetic structure of S. kurtzianum populations, and discuss the importance of field studies on population genetics, reproductive biology and ecology to design collections and conservation strategies.     

Elevated Mutagenicity in Meiosis and Its Mechanism

Diploid germ cells produce haploid gametes through meiosis, a unique type of cell division. Independent reassortment of parental chromosomes and their recombination leads to ample genetic variability among the gametes. Importantly, new mutations also occur during meiosis, at frequencies much higher than during the mitotic cell cycles. These meiotic mutations are associated with genetic recombination and depend on double‐strand breaks (DSBs) that initiate crossing over. Indeed, sequence variation among related strains is greater around recombination hotspots than elsewhere in the genome, presumably resulting from recombination‐associated mutations. Significantly, enhanced mutagenicity in meiosis may lead to faster divergence during evolution, as germ‐line mutations are the ones that are transmitted to the progeny and thus have an evolutionary impact. The molecular basis for mutagenicity in meiosis may be related to the repair of meiotic DSBs by polymerases, or to the exposure of single‐strand DNA to mutagenic agents during its repair.