Anther culture for haploid and doubled haploid production

Haploids are plants with a gametophytic chromosome number and doubled haploids are haploids that have undergone chromosome duplication. The production of haploids and doubled haploids (DHs) through gametic embryogenesis allows a single-step development of complete homozygous lines from heterozygous parents, shortening the time required to produce homozygous plants in comparison with the conventional breeding methods that employ several generations of selfing. The production of haploids and DHs provides a particularly attractive biotechnological tool, and the development of haploidy technology and protocols to produce homozygous plants has had a significant impact on agricultural systems. Nowadays, these biotechnologies represent an integral part of the breeding programmes of many agronomically important crops. There are several available methods to obtain haploids and DHs, of which in vitro anther or isolated microspore culture are the most effective and widely used. This review article deals with the current status of knowledge on the production of haploids and DHs through pollen embryogenesis and, in particular, anther culture.     

Gametic embryogenesis and haploid technology as valuable support to plant breeding

Plant breeding is focused on continuously increasing crop production to meet the needs of an ever-growing world population, improving food quality to ensure a long and healthy life and address the problems of global warming and environment pollution, together with the challenges of developing novel sources of biofuels. The breeders’ search for novel genetic combinations, with which to select plants with improved traits to satisfy both farmers and consumers, is endless. About half of the dramatic increase in crop yield obtained in the second half of the last century has been achieved thanks to the results of genetic improvement, while the residual advance has been due to the enhanced management techniques (pest and disease control, fertilization, and irrigation). Biotechnologies provide powerful tools for plant breeding, and among these ones, tissue culture, particularly haploid and doubled haploid technology, can effectively help to select superior plants. In fact, haploids (Hs), which are plants with gametophytic chromosome number, and doubled haploids (DHs), which are haploids that have undergone chromosome duplication, represent a particularly attractive biotechnological method to accelerate plant breeding. Currently, haploid technology, making possible through gametic embryogenesis the single-step development of complete homozygous lines from heterozygous parents, has already had a huge impact on agricultural systems of many agronomically important crops, representing an integral part in their improvement programmes. The aim of this review was to provide some background, recent advances, and future prospective on the employment of haploid technology through gametic embryogenesis as a powerful tool to support plant breeding.     

Isopropyl-N(3-chlorophenyl) carbamate (CIPC) induced chromosomal loss in soybean: a new tool for plant somatic cell genetics

Summary The properties and uses of tissue culture partial haploid soybean cell lines are explored. Partial haploid lines were prepared by CIPC treatment of a genetic heterozygote, and compared to this heterozygote and to homozygous cell lines corresponding to the parental genotypes from which the heterozygote was derived. Cell lines which lack chromosomes were characterized physiologically and with respect to a variety of isozyme markers. Often the loss of chromosomes revealed a phenotype corresponding to a recessive parental genotype. In some cases, however, new phenotypes were observed indicating a complex genotype and suggests the interaction of several genes. The implications of this for plant breeding are discussed.CIPC also was used as a tool to dissect a complex phenotype which arose as the result of mutagenesis. A mutant cell line which required asparagine for growth but also had acquired the ability to grow on allantoin as a sole source of nitrogen was treated with CIPC to remove chromosomes. The requirement for asparagine could be separated from the ability to use allantoin, demonstrating that these phenotypes were the result of separate mutations.     

Inheritance patterns of the response to in vitro doubled haploid induction in perennial ryegrass (<Emphasis Type="Italic">Lolium perenne</Emphasis> L.)

The ability to produce doubled haploid (DH) plants has found broad application in research and breeding. For major crop species such as maize (Zea mays L.) and barley (Hordeum vulgare L.), routine large-scale production of DHs has enabled the acceleration of breeding processes, for example through efficient generation of homozygous lines. However, in forage crops such as perennial ryegrass (Lolium perenne L.), low and genotype-specific responses to in vitro anther culture (AC) still limit wide-spread use of DHs. Here, we report the responses of nine bi-parental populations, segregating for microspore embryogenesis and plant regeneration capacity, to an effective AC protocol. Genotypes of exceptionally high androgenic ability, producing over 200 green plants per 100 anthers cultured, could be selected. Continuous and distinctly shaped distributions for the evaluated traits were indicative of quantitative polygenic control and the presence of different alleles in each population. An insignificant association of embryo production with plant regeneration, as well as a low correlation between green and albino plant yield (ρ?=?0.20), suggested that different genes influence these traits. The populations evaluated here provide a rich source of alleles needed for the introgression of high levels of androgenic capacity into recalcitrant material. Moreover, this germplasm is ideally suited for use in future genotyping and mapping studies so that the genetic control of androgenic capacity in perennial ryegrass can be elucidated. Ultimately, our results will help to realize the potential of DH induction in one of the world’s most important forage crop species.     

Genetic and physiological variability among cultured cells and regenerated plants of Nicotiana tabacum

Summary Tobacco cell lines selected for resistance to picloram (4-amino-3,5,6-trichloropicolinic acid) and plants regenerated from these cell lines manifest several traits not shown by the parental strains. Genetics analyses of the regenerated plants have permitted the sources of this variability to be identified.Tricotyledenous seedlings appeared at a much higher frequency among the progeny of a heterozygous mutant plant (PmR1/+) regenerated from culture than they did among progeny of normal regenerated plants. In crosses with the regenerated heterozygous mutant plant and with homozygous progeny of this plant (PmR1/PmR1) the frequency of tricotyly was influenced more by the generation than by the genotype of the parent plant. Therefore, it is concluded that tricotyly is a physiological response to passage through cell culture.More than half of the picloram-resistant cell lines isolated were also resistant to hydroxyurea. Segregation of these two resistances was analyzed in progeny of crosses with regenerated plants. In all cases hydroxyurea-resistance was genetically stable and inherited as a single dominant nuclear mutation (designated HuR). In crosses with plants PmR1/+ and PmR7/+ the HuR and PmR mutations assorted independently. In contrast, the HuR mutation recovered from plant PmR6/+ was linked to the PmR6 mutation.     

Single large-scale marker-assisted selection (SLS-MAS)

This paper presents a new approach for plant improvement that interactively combines the use of DNA markers and conventional breeding. This approach involves selecting plants at early generation with a fixed, favorable genetic background at specific loci, conducting a single large-scale marker-assisted selection (SLS-MAS) while maintaining as much as possible the allelic segregation in the rest of the genome. First, the identification of elite lines presenting high allelic complementarity and being outstanding for traits of interest is required to capture favorable alleles from different parental lines. Second, after identification of the most favorable genomic regions for each selected parental line, those lines are intercrossed to develop segregating populations from which plants homozygous for favorable alleles at target loci are selected. One objective of the scheme is to conduct the marker-assisted selection only once, and it requires the selection of a minimum number of plants to maintain sufficient allelic variability at the unselected loci. Therefore, the selection pressure exerted on the segregating population is quite high and the screening of large populations is required to achieve the objectives of the scheme. No selection is applied outside the target genomic regions, to maintain as much as possible the Mendelian allelic segregation among the selected genotypes. After selection with DNA markers, the genetic diversity at un-selected loci may allow breeders to generate new varieties and hybrids through conventional breeding in response to various local needs. Although the single large-scale MAS scheme described here is oriented toward maize and large-scale breeding programs with substantial resources, the flexibility of this scheme would allow breeding programs to develop options compatible with local resources.     

Marker-assisted selection for low phytic acid (lpa1-1) with single nucleotide polymorphism marker and amplified fragment length polymorphisms for background selection in a maize backcross breeding programme

The level of phytic acid is difficult to assess in a maize breeding programme, therefore a co-dominant single nucleotide polymorphism (SNP) marker was used to detect the single recessive low phytic acid (lpa1-1) gene in a BC2F1 population developed from a locally adapted tropical normal inbred line (P 16) and CM 32 (lpa1-1 donor). High-resolution melt analysis of the lpa1-1 SNP marker was able to identify 11 homozygous recessive and 17 heterozygote genotypes for the lpa1-1 mutation. The SNP R ² values for the heterozygotes were higher (90.95?C99.59%) than the lpa1-1 recessives (82.81?C99.58%). The selected BC2F1 lines were fingerprinted with six amplified fragment length polymorphism (AFLP) EcoRI/MseI primer combinations to determine the amount of recurrent parent genome present. The 277 AFLP markers were clearly able to differentiate all the BC2F1 lines from each other and the parental controls with a similarity range from 62.12 to 92.15%. It is expected in the BC2 generation to find 87.5% similarity to the recurrent parent, however in this study higher levels of similarity in 13 BC2F1 lines (six heterozygotes and seven homozygous recessive) with 92.15?C83.33% similarity were observed. The use of marker-assisted selection for foreground and background selection greatly increased the efficiency of detection of the homozygous recessive (99.58%) and heterozygous (99.59%) genotypes as well as improving the recovery of the recurrent parent (92.15%) in the BC2F1 generation of the maize backcross breeding programme.     

水稻抗草品种选育研究初报

利用PI312777抗草稻种材料,与本单位优质稻核心种质及其衍生系统材料广泛杂交配组,创建一批以PI312777衍生系统为主的新型育种材料。建立水稻抗草育种圃,重点针对抗草特性和综合农艺性状,通过田间抑草效应筛选,培育水稻抗草苗头株系,在稳定世代(F4代以上)应用以水稻特征次生物质为标记评价水稻品种(系)化感潜力的方法,辅助选育水稻抗草新品种(系)。初步提出适合华南稻区的以田间筛选和评价为主,以特征次生物质标记评价水稻品种(系)化感潜力的方法为辅,田间与实验室相结合,操作性强、简便有效的水稻抗草育种方法。     

Cassava breeding: opportunities and challenges

Although cassava is a major food crop, its scientific breeding began only recently compared with other crops. Significant progress has been achieved, particularly in Asia where cassava is used mainly for industrial processes and no major biotic constraints affect its productivity. Cassava breeding faces several limitations that need to be addressed. The heterozygous nature of the crop and parental lines used to generate new segregating progenies makes it difficult to identify parents with good breeding values. Breeding so far has been mainly based on a mass phenotypic recurrent selection. There is very little knowledge on the inheritance of traits of agronomic relevance. Several approaches have been taken to overcome the constraints in the current methodologies for the genetic improvement of cassava. Evaluations at early stages of selection allow for estimates of general combining ability effect or breeding values of parental lines. Inbreeding by sequential self-pollination facilitates the identification of useful recessive traits, either already present in the Manihot gene pool or induced by mutagenesis.