玉米单倍体加倍方法研究进展

主要综述了近年来玉米单倍体加倍方法的最新研究进展。自然加倍法和以秋水仙素及除草剂为加倍剂的化学加倍法是人们研究较多的加倍方法。     

The effect of spikelet position on the success frequencies of wheat haploid production using the maize cross system

Barbela is an old Portuguese landrace of wheat that is highly genetically heterogeneous. Different Barbela populations when subjected to aluminium stress show variable levels of tolerance. In order to study the inheritance of this character, doubled haploid (DH) lines were developed. These DH were obtained by intergeneric crosses of 14 different lines of Barbela with maize. During this process the efficiency of the technique was evaluated and suggestions for its improvement were obtained. Several parameters were studied in the crosses: % of crossability, % of embryos per florets pollinated and % of embryos per seed set. The different genotypes of Barbela showed significant variation for the parameters analysed. When the reciprocal crosses were analysed, no differences were found, indicating that cytoplasm differences do not influence the parameters of DH production. However, different spikelet positions (lower, middle and upper) gave highly significant differences in all parameters analysed. Highest success frequencies were obtained for pollinated spikelets in the middle of the spike. This can indicate that concentrating on the middle part of the spike can increase the frequency of DHs obtained using inter generic crosses of wheat with maize. This revised version was published online in August 2006 with corrections to the Cover Date.     

Detecting and estimating segregation distortion and linkage between glufosinate tolerance and blackleg resistance in Brassica napus L.

Summary The segregation and linkage between glufosinate (transgenes ‘Rf3’ and ‘T177’) and blackleg resistance genes in canola (Brassica napus L.) were assessed using F₁ microspore-derived doubled haploid (DH) populations from four crosses including reciprocals, two involving the transgene ‘Rf3’ and the other two involving the transgene ‘T177’. To relax the assumption of no segregation distortion required for the conventional analysis of segregation and linkage, we employed Bailey's analysis that allows detecting segregation distortion at linked loci. The significant departures from the 1:1 segregation were detected in the crosses involving the transgene ‘T177’ but not in the crosses involving the transgene ‘Rf3’. The apparent deficit of the herbicide tolerant DH lines in the crosses with the transgene ‘T177’ is likely due to differential selection against the gametes carrying ‘T177’ during microspore culture. The linkage was strong between blackleg resistance and the transgene ‘Rf3’ but weak or absent between blackleg resistance and the transgene ‘T177’, suggesting that the two transgenes are probably inserted into distant regions of the genome. The observed linkage offers an opportunity to develop new canola cultivars with both glufosinate tolerance conferred by transgene ‘Rf3’ and blackleg resistance.     

Enhanced anther culture efficiency of indica rice (Oryza sativa L.) through modification of the culture media

The production of microspore-derived green plants from anther culture of indica rice is generally very low compared with japonica cultivars. A modified anther culture medium, consisting of a higher KNO₃ content (31 mm ) and casein hydrolysate (CH, 500 mg/1) but without ammonium salts, was tested in comparison with a medium consisting of the widely-used N6 medium nitrogen background, using four indica × indica F₁ hybrids as test materials. Green plant regeneration frequency was at least three-fold higher in the microspore-calli derived from the former medium than in those derived from the modified N6 medium. More than 700 microspore-derived plants were raised in the field. Another study was carried out using indica × japonica and indica × javanica F₁ hybrids. The results indicated that a medium with higher (3.5 mm ) ammonium sulphate may induce a higher frequency of anthers with microspore-calli but not necessarily lead to a larger number of green-plant regenerating calli. Subsequently, using the indica cv. ‘IR-43’ as the test material, use of a lower level (1.75 mm ) of (NH₄)₂SO₄, in addition to KNO₃ (31 mm ), was found to be better than CH (500 mg/l) for anther-response as well as green plant regenerability of the derived microspore-calli. Nitrate-nitrogen or ammonium-nitrogen alone elicited poor response. Twenty-five media involving combinations of KNO₃ (20–34 mm ) and (NH₄)₂SO₄ (1–3 mm ) were tested for their effects on anther response. Combinations involving KNO₃ (31–34 mM) and (NH₄)₂SO₄ (2.0–2.5 mm ) were found superior not only for achieving greater anther response but also, for subsequent green-plant regeneration. This contrasts with the 28 mm of KNO₃ and 3.5 mm of (NH₄)₂SO₄ in the widely-used N6 medium developed for japonica rice. Other than potassium nitrate and ammonium sulphate, and potassium phosphate to some extent, the levels of other inorganic salts tested did not make any significant difference to the process of anther response. Based on these results, modified media with three levels of ammonium sulphate were tested for anther culture efficiency of indica × japonica and indica × javanica derivatives (F₃s). Microspore-calli derived from a medium of a lower (1.75 mm ) level of (NH₄)₂SO₄ showed a higher regeneration potential overall than those derived from a higher (3.5 mm ) level. A revised medium has been suggested, on the basis of these results, for the realization of improved anther culture efficiency and, consequently, improved feasibility of using doubled haploids in genetic and breeding research with indica rice.     

Genetics of multiple disease resistance in a doubled-haploid population of barley

The barley accession Q21861 possesses resistance to the stem-rust (Puccinia graminis f.sp. tritici), leaf-rust (P. hordei), and powdery-mildew (Blumeria graminis f.sp. hordei) pathogens. An anther-culture-derived doubled-haploid population was produced from F₁ plants from a cross of this accession and the susceptible breeding line SM89010 as a means of rapidly and efficiently determining the genetics of multiple disease resistance. The doubled-haploid population segregated 1:1 (resistant:susceptible) for resistance to the stem rust pathotype QCC indicating the involvement of a single resistance gene, rpg4. Two-gene (3:1) and one-gene (1:1) segregation ratios were observed for resistance to the stem-rust pathotype MCC at low (23–25°c) and high (27–29°C) temperature, respectively. These different segregation patterns were due to a pathotype × temperature interaction exhibited by rpg4 and Rpg1. another stem-rust-resistance gene present in Q21861. One-gene and two-gene segregation ratios were observed in reaction to the leaf rust and powdery mildew pathogens. These data demonstrate the utility of doubled haploid populations for determining the genetics of multiple disease resistance in barley.     

辣椒单倍体与双单倍体植株叶片光合作用研究

对辣椒单倍体和双单倍体植株的光合作用进行了研究。结果表明 ,在不同光照条件下 ,单倍体植株净光合速率均明显比双单倍体低。在弱光照下 ,气孔的限制作用是单倍体植株净光合速率较低的主要原因 ;而在正常光照下 ,非气孔限制因素是净光合速率降低的主要原因。单倍体植株在强光下受到明显的光抑制 ,单倍体植株净光合速率、气孔导度和表观量子效率均较低 ,在正常光照条件下羧化效率较低 ;光补偿点和细胞间隙CO2 补偿点较高等均是单倍体植株的重要特征     

Analysis of durable resistance to stem rust in barley

Summary Since the mid-1940's, barley cultivars grown in the northern Great Plains of the USA and Canada have been resistant to stem rust caused byPuccinia graminis f. sp.tritici. This durable resistance is largely conferred by a single gene,Rpg1, derived from a single plant selection of the cultivar Wisconsin 37 and an unimproved Swiss cultivar. At the seedling stage, barley genotypes withRpg1 generally exhibit low mesothetic reactions at 16–20° C and slightly higher mesothetic reactions at 24–28° C to many stem rust pathotypes. This resistance is manifested by a low level of rust infection and mostly incompatible type uredia on adult plants.Rpg1 reacts in a pathotype-specific manner since some genotypes ofP. g. f. sp.tritici are virulent on cultivars carrying this gene in the field. Several factors may have contributed to the longevity of stem rust resistance in barley, a) since barley is planted early and matures early, it can sometimes escape damage from stem rust inoculum carried from the south; b) one or more minor genes may augment the level of resistance already provided byRpg1; c) the cultivation of resistant wheat cultivars and eradication of barberry have reduced the effective population size and number of potential new pathotypes ofP. g. f. sp.tritici, respectively; and d) virulent pathotypes ofP. g. f. sp.tritici andP. g. f. sp.secalis have not become established. This situation changed in 1989 when a virulent pathotype (Pgt-QCC) ofP. g. f. sp.tritici became widely distributed over the Great Plains. However,Rpg1 may still confer some degree of resistance to pathotype QCC because stem rust severities have been low to moderate and yield losses light on barley cultivars carrying the gene during the last four seasons (1989–1992). Several sources of incomplete resistance to pathotype QCC have been identified in barley. To facilitate the transfer of resistance genes from these sources into advanced breeding lines, molecular marker assisted selection is being employed.     

Production of Haploid Sugar Beet (Beta vulgaris L.) by Culturing Unpollinated Ovules

The culture of unpollinated ovules is shown to be a suitable system for the production of haploid sugar beet (Beta vulgaris L.). The yield of haploids depended upon the genotype and varied between 0 and 13 % with a mean of 1.0 %. Haploid plants could be produced from approximately 50 % of all genotypes examined. The majority of the haploids isolated (about 90%) maintained the haploid genome level during the in vitro culture and propagation; 10% of the haploid clones showed a spontaneous doubling to the diploid genome level.     

Analysis of anther culture response in hexaploid triticale

The androgenic response of five varieties and 10 F₁ intervarietal hybrids of 6 × triticale was analysed. The overall response involved three independent phenomena: the production of embryoids, the capability of embryoids to regenerate plants, and the production of green plants. Each of these factors appeared to be controlled by independent genetic systems. From a practical point of view, the product of these three factors, the number of green haploid plants per anther (GPA), is the most reliable variable for the evaluation of genotypes. Positive heterotic effects on GPA were observed in the hybrids, which may indicate that GPA is controlled predominantly by additive genes. No correlation was found between the number of green haploid plants produced by hybrids and that yielded by the best parental line. Therefore, it is difficult to predict the yield of green haploid plants that can be produced by a hybrid simply from the number produced by its parents.     

Somatic embryogenesis from floral tissue of cassava (Manihot esculenta Crantz)

The aim of this study was to examine the embryogenic potential of floral material of the cassava cultivar MCOL 1505. Macerated immature inflorescences were found to be highly embryogenic, with almost 78% of the explants producing somatic embryos. Somatic embryos were also produced from whole male florets and half florets although at much lower rates. No regeneration was obtained from anther, microspore or floret wall tissue. Somatic embryos derived from immature inflorescences were regenerated via organogenesis and the plants derived from this process were assessed in terms of phenotype and ploidy level. If haploid plants could be produced by this method, this would have significant implications in assisting traditional cassava breeding, as this would allow homozygosity to be reached more rapidly. In a crop such as cassava, which is highly heterozygous in nature, the use of haploids in a breeding programme could considerably shorten the time taken to produce new desirable cultivars. This is the first report on plant regeneration through somatic embryogenesis from floral tissue of cassava. This revised version was published online in July 2006 with corrections to the Cover Date.