Sang Woo Bang

Structural and Biochemical Dissection of Photorespiration in Hybrids Differing in Genome Constitution between Diplotaxis tenuifolia (C3-C4) and Radish (C3)

We compared the structural, biochemical, and physiological characteristics involved in photorespiration of intergeneric hybrids differing in genome constitution (DtDtR, DtDtRR, and DtRR) between the C3-C4 intermediate species Diplotaxis tenuifolia (DtDt) and the C3 species radish (Raphanus sativus; RR). The bundle sheath (BS) cells in D. tenuifolia included many centripetally located chloroplasts and mitochondria, but those of radish had only a few chloroplasts and mitochondria. In the hybrids, the numbers of chloroplasts and mitochondria, the ratio of centripetally located organelles to total organelles, and the mitochondrial size in the BS cells increased with an increase in the constitution ratio of the Dt:R genome. The P-protein of glycine decarboxylase (GDC) was confined to the BS mitochondria in D. tenuifolia, whereas in radish, it accumulated more densely in the mesophyll than in the BS mitochondria. In the hybrids, more intense accumulation of GDC in the BS relative to the mesophyll mitochondria occurred with an increase in the Dt:R ratio. These structural and biochemical features in the hybrids were reflected in the gas exchange characteristics of leaves, such as the CO2 compensation point. Our data indicate that the leaf structure, the intercellular pattern of GDC expression, and the gas exchange characteristics of C3-C4 intermediate photosynthesis are inherited in the hybrids depending on the constitution ratio of the parent genomes. Our findings also demonstrate that the apparent reduced photorespiration in C3-C4 intermediate plants is mainly due to the structural differentiation of mitochondria and chloroplasts in the BS cells combined with the BS-dominant expression of GDC.

Interspecific and intergeneric hybridization and chromosomal engineering of Brassicaceae crops

In Brassicaceae crop breeding programs, wild relatives have been evaluated as genetic resources to develop new cultivars with biotic and abiotic stress resistance. This has become necessary because of the diversification of ecotypes of diseases and pests, changing food preferences, advances in production technology, the use of new approaches such as in vitro breeding programs, and the need for economical production of F1 seed. To produce potential new cultivars, interspecific and intergeneric hybridizations have been performed between cultivated species and between cultivated species and their wild relatives. Furthermore, interspecific and intergeneric hybrids have been successfully produced using embryo rescue techniques. In this paper, we review the interspecific and intergeneric incompatibilities between Brassicaceae crops and their wild relatives, and the production, characterization, and improvement of synthetic amphidiploid lines, alien gene introgression lines, alloplasmic lines, monosomic alien chromosome addition lines, and monosomic alien chromosome substitution lines. The goal is to provide useful materials to support practical breeding strategies and to study the genetic effects of individual chromosomes on plant traits, the number of genes that control a trait, their linkage relationships, and genetic improvement in Brassicaceae crops.

Genetic analysis of hybrid seed formation ability of Brassica rapa in intergeneric crossings with Raphanus sativus

A hybridization barrier leads to the inability of seed formation after intergeneric crossings between Brassica rapa and Raphanus sativus. Most B. rapa lines cannot set intergeneric hybrid seeds because of embryo breakdown, but a B. rapa line obtained from turnip cultivar ‘Shogoin-kabu’ is able to produce a large number of hybrid seeds as a maternal parent by crossings with R. sativus. In ‘Shogoin-kabu’ crossed with R. sativus, developments of embryos and endosperms were slower than those in intraspecific crossings, but some of them grew to mature seeds without embryo breakdown. Intergeneric hybrid seeds were obtained in a ‘Shogoin-kabu’ line at a rate of 0.13 per pollinated flower, while no hybrid seeds were obtained in a line developed from Chinese cabbage cultivar ‘Chiifu’. F1 hybrid plants between the lines of ‘Shogoin-kabu’ and ‘Chiifu’ set a larger number of hybrid seeds per flower, 0.68, than both the parental lines. Quantitative trait loci (QTLs) for hybrid seed formation were analyzed after intergeneric crossings using two different F2 populations derived from the F1 hybrids, and three QTLs with significant logarithm of odds scores were detected. Among them, two QTLs, i.e., one in linkage group A10 and the other in linkage group A01, were detected in both the F2 populations. These two QTLs had contrary effects on the number of hybrid seeds. Epistatic interaction between these two QTLs was revealed. Possible candidate genes controlling hybrid seed formation ability in QTL regions were inferred using the published B. rapa genome sequences.

Production and characterization of an alloplasmic and monosomic addition line of Brassica rapa carrying the cytoplasm and one chromosome of Moricandia arvensis.

Intergeneric hybridization was performed between Moricandia arvensis and four inbred lines of Brassica rapa following embryo rescue. Three F1 hybrid plants were developed from three cross combinations of M. arvensis × B. rapa, and amphidiploids were synthesized by colchicine treatment. Six BC1 plants were generated from a single cross combination of amphidipolid × B. rapa ‘Ko1-303’ through embryo rescue. One BC2 and three BC3 plants were obtained from successive backcrossing with B. rapa ‘Ko1-303’ employing embryo rescue. Alloplasmic and monosomic addition lines of B. rapa (Allo-MALs, 2n = 21) were obtained from backcrossed progeny of three BC3 plants (2n = 21, 22 and 23) without embryo rescue. An alloplasmic line of B. rapa (2n = 20) degenerated before floliation on 1/2 MS medium due to severe chlorosis. Allo-MALs of B. rapa (2n = 21) showed stable male sterility without any abnormal traits in vegetative growth and female fertility. Molecular analyses revealed that the same chromosome and cytoplasm of M. arvensis had been added to each Allo-MAL of B. rapa. This Allo-MAL of B. rapa may be useful material for producing cytoplasmic male sterile lines of B. rapa.

Inheritance of C3-C4 Intermediate Photosynthesis in Reciprocal Hybrids between Moricandia arvensis (C3-C4) and Brassica oleracea (C3) that Differ in their Genome Constitution

Abstract To Elucidate The Genetic Mechanisms Underlying C3―C4 intermediate Photosynthesis, We investigated The Structural and Photosynthetic Characteristics of Leaves of Reciprocal Hybrids Between The C3―C4 intermediate Species Moricandia Arvensis (L.) Dc. (Mama) and The C3 Species Brassica Oleracea L. (Cabbage; Cc), Which Differ in Genome Constitution. Moricandia Arvensis Bundle Sheath (Bs) Cells included Many Centripetally Located Chloroplasts and Mitochondria, Whereas Those of Cabbage Had Few Organelles. Hybrid Leaves Were Structurally intermediate Between Those of The Parents and Showed Stronger intermediate C3―C4 Features As The Proportion of The Ma Genome increased. The P-Protein of Glycine Decarboxylase (Gdc) Was Confined Mainly To Bs Mitochondria in M. Arvensis, But Accumulated More in The Mesophyll (M) of Cabbage. in The Hybrids, The Accumulation of Gdc in Bs Cells increased With An increasing Ma:C Ratio. Hybrids Exhibited Gradients in Structural and Biochemical Features, Even in Reciprocal Crosses. The Co2 Compensation Point of Reciprocal Hybrids With High Ma:C Ratios Was Lower Than That of Cabbage But Higher Than That of M. Arvensis. Thus, The Structural and Biochemical Features in Hybrid Leaves Reduced Photorespiration. Moricandia Arvensis Had A Higher Photosynthetic Rate Than Cabbage, But The Photosynthetic Rates of Hybrids Were intermediate Between Those of The Parents Or Comparable To That of M. Arvensis. Our Results Demonstrate That The C3―C4 intermediate Characteristics Are inherited Based On The Ratio of The Parent Genomes, and That There Is No Evidence of Cytoplasmic inheritance in These Characteristics.

The use of species-specific DNA markers for assessing alien chromosome transfer in Brassica rapa and Brassica oleracea-monosomic additions of Raphanus sativus

Abstract Monosomic addition lines (MALs) are useful materials not only for cytogenetic and molecular genetic studies but also for plant breeding as gene sources. In our previous study, two MALs in the tribe Brassiceae were developed, one being Raphanus sativus lines with alien chromosomes of Brassica rapa (B. rapa-monosomic addition lines; BrMALs) and the second being those with alien chromosomes of Brassica oleracea (B. oleracea-monosomic addition lines; BoMALs). We developed species-specific DNA markers from the genomic sequences of B. rapa and B. oleracea comparing them with those of R. sativus, and identified chromosomes added in BrMALs and BoMALs using these markers. It was revealed that eight types of BrMALs have seven chromosomes of B. rapa and seven types of BoMALs have six chromosomes of B. oleracea. Furthermore, chromosome breakage and homoeologous recombination were suggested to have occurred in some MALs. The developed species-specific DNA markers are considered to be useful for producing MALs and also for assessing chromosome abnormality in MALs.

The Use of Genome Information for Intergeneric Hybridization Breeding

Radish is an important vegetable crop and has several agriculturally useful traits to improve the breeding of Brassica crops, which include cytoplasmic male sterility, resistance to pod shattering, tolerance to saline soils, and resistance to nematode and diseases. To transfer these traits into Brassica, a number of intergeneric hybridization have been performed using intergeneric cross and/or protoplast fusion, and it was shown that some of them were introduced successfully into and expressed in Brassica or hybrids with Brassica. The various kinds of hybrid progenies such as synthetic amphidiploid line, alloplasmic line, alien gene(s) introgression line, and monosomic alien chromosome addition line would be valuable genetic resources not only to breed more useful cultivars but also to analyze each chromosome and gene concerned. Recently, the development of numerous molecular markers for genetic study has been accelerated in radish, and a high-density linkage map has been constructed. These molecular tools have been used for the identification of QTLs associated with important agronomic traits of radish, genetic analysis of hybridization barrier between radish and Brassica crops, and comparative genomics of Brassicaceae. The further accumulation of genome information will facilitate the cloning of candidate genes in QTLs and the marker-assisted breeding in radish, in addition to the intergenomic transfer of useful radish genes into Brassica crops through intergeneric hybridization.