香蕉原生质体培养和体细胞杂交研究进展

香蕉是继水稻、小麦、玉米之后的世界第4大粮食作物和近5亿发展中国家人口的主要粮食,香蕉产业的健康发展具有重要的经济和社会地位。但由于香蕉品种的高度不育性和多倍性,制约了用传统育种方法培育生产实践中所需抗病虫害新品种。通过原生质体融合进行的体细胞杂交育种有望克服这些障碍。原生质体培养技术是细胞融合所必需的基础技术之一,自1984年首次进行香蕉原生质体分离研究以来,有关香蕉原生质体分离、培养和融合等方面的研究陆续开展,相继有超过10个品种成功获得再生植株,并且开始了体细胞杂交研究的探索。作者就香蕉原生质体培养和体细胞杂交研究进展进行综述。     

抗寒甘蓝型油菜的初步选育

为将白菜型油菜(Brassica rapa L.)中的抗寒性状转育到甘蓝型油菜(Brassica napus L.)中,以3个甘蓝型油菜品系为母本,4个抗寒白菜型油菜品种为父本配制杂交组合,研究了各杂交组合回交亲和性以及杂种的形态表现.结果表明:①甘白种间杂交亲和性较高,亲和指数最高的组合达3.42,最低为0.50;②杂交亲和性受双亲基因型的影响,且母本基因型对杂交亲和指教的影响大于父本基因型;③利用SSR鉴定真假杂种,真杂种叶片形态特征介于双亲之间,根系形态偏向于白菜型油菜,具有粗壮的主根而须根较少.通过测配可选出杂交亲和性较高的甘白组合,从而繁殖出较多的杂交及回交分离后代用于下一步的田间抗寒性鉴定和筛选.     

Effects of marigold (Tagetes sp.) roots on soil microorganisms

 Marigolds (genus Tagetes) suppress populations of soil endopathogenic nematodes such as Pratylenchus penetrans and Meloidogyne species. Nematode suppression by marigolds is thought to be due to thiophenes, heterocyclic sulfur-containing molecules abundant in this plant. When activated, thiophenes such as α-terthienyl produce oxygen radicals. If marigold roots release such a powerful biocidal agent and it is activated in soil, microbial populations in the marigold rhizosphere should be substantially perturbed. We made various measurements of microbial population size and activity in soils that had been cropped to marigolds (Crackerjack, Creole) in the field and in the greenhouse, and compared these with bare soil and soil cropped to rye (Secale cereale L.). Total extractable microbial biomass (measured by the fumigation extraction method), total bacteria (measured by epifluorescence microscopy on 5-(4,6-dichlorotriazine-2-γl) aminofluorescein-stained preparations), heterotrophic bacteria (measured by plate count on various media), and nitrite-oxidizing bacteria (measured by the most-probable-number method) were not significantly different in any of the treatments. Residues of ¹⁴C-labelled rye were mineralized slightly more rapidly in rye-cropped soil than in the other treatments, which were comparable. The rates of die-back of introduced cells of the bacteria Escherichia coli and Rhodococcus TE1 were similar in marigold-cropped and control soils, suggesting that there was not a noteworthy accumulation of biocidal agents in soils cropped to marigolds. We conclude that marigolds do not cause a general depression in the numbers of microorganisms in soils, and that nematode control by this plant may not be due to the release of a biocidal agent into the soil. Received: 4 September 1997     

Embryo development in two barley genotypes after self pollination and pollination with Hordeum bulbosum L.

Summary Embryo development in vivo was compared in the two barley genotypes VK 16032 and Vogelsanger Gold after self pollination and after pollination with H. bulbosum. Embryo growth in VK 16032 after crossing with H. bulbosum showed a similar growth pattern to that of the self pollinated embryos, although the size increase was smaller. The hybrid embryos continued to grow until day 18 after pollination. The embryos from the cross between Vogelsanger Gold and H. bulbosum ceased to grow 8 days after pollination. Arrested embryo growth with subsequent abortion in Vogelsanger Gold was associated with a very early depletion and break down of the endosperm. Use of barley genotypes less sensitive to the genomic disturbances after crossing with H. bulbosum in combination with genotypes like Vogelsanger Gold is recommended in haploid breeding work.     

Diversity of Phytophthora species in natural ecosystems of Taiwan and association with disease symptoms

In 2013 a survey of Phytophthora diversity was performed in 25 natural and seminatural forest stands and 25 rivers in temperate montane and subtropical lowland regions of Taiwan. Using baiting assays, 10 described species and 17 previously unknown taxa of Phytophthora were isolated from 71.5% of the 144 rhizosphere soil samples from 33 of 40 tree species sampled in 24 forest stands, and from 19 rivers: P. capensis, P. citrophthora, P. plurivora, P. tropicalis, P. citricola VII, P. sp. × botryosa‐like, P. sp. × meadii‐like and P. sp. occultans‐like from Clade 2; P. palmivora from Clade 4; P. castaneae and P. heveae from Clade 5; P. chlamydospora and P. sp. forestsoil‐like from Clade 6; P. cinnamomi (Pc), P. parvispora, P. attenuata nom. prov., P. flexuosa nom. prov., P. formosa nom. prov., P. intricata nom. prov., P. × incrassata nom. prov. and P. × heterohybrida nom. prov. from Clade 7; P. sp. palustris and five new hybrid species from Clade 9. The A1 mating type of Pc was widespread in both montane and lowland forests and rarely associated with disease, whereas the A2 mating type was limited to lowland forests and in some cases causing severe dieback. Most other Phytophthora species were not associated with obvious disease symptoms. It is concluded that (i) Taiwan is within the centre of origin of most Phytophthora taxa found, (ii) Pc A2 is an introduced invasive pathogen, and (iii) interspecific hybridizations play a major role in speciation and species radiations in diverse natural ecosystems.     

Occurrence and diversity of Tomato spotted wilt virus isolates breaking the Tsw resistance gene of Capsicum chinense in Yunnan,southwest China

Widely used resistant peppers (Capsicum spp.) bearing the Tsw locus triggered the rapid emergence of resistance‐breaking (RB) isolates of Tomato spotted wilt virus (TSWV) around the world. However, although TSWV‐induced diseases have rapidly increased in Yunnan, southwest China, in recent years, no information is available about the diversity of TSWV isolates in this region. In this study, the occurrence of natural TSWV RB variants among isolates collected in Yunnan is reported. Initially, a TSWV isolate from asparagus lettuce (TSWV‐LE) was collected in Yunnan in 2012. Surprisingly, this isolate of TSWV induced systemic necrosis on pepper carrying the Tsw resistance gene. Novel TSWV isolates, collected in 2015, included a tomato isolate (TSWV‐YN18) and a tobacco isolate (TSWV‐YN53) that also overcame Tsw‐mediated resistance. TSWV‐YN18 induced systemic ringspots, whereas TSWV‐YN53 caused systemic chlorotic mottling. Variations in the TSWV nonstructural (NSs) protein are the key determinants associated with Tsw resistance‐breaking isolates. It was found that TSWV‐LE NSs retained the hypersensitive response (HR) induction, whereas TSWV‐YN18 and TSWV‐YN53 NSs were unable to induce HR. However, the NSs of all three RB isolates suppressed RNA silencing. Sequence analysis of the NSs revealed that RB isolates of Yunnan have no amino acid mutation sites common to other previously reported RB isolates. However, two amino acids (F74 and K272) on TSWV‐LE NSs make it distinct from TSWV‐YN18 and TSWV‐YN53. The occurrence of different RB isolates and the failure of Tsw‐mediated resistance control pose serious threats to domestic pepper crops in southwest China.     

The development of a rapid PCR assay for detection of Fusarium moniliforme

The fungus Fusarium moniliforme infects a wide range of crops throughout the world. In maize (Zea mays L.) it causes seedling blight and root, stalk, and ear rots. A simple procedure that can be used to detect infection by F. moliliforme from infected plant tissues has been developed. A F. moniliforme genomic library was prepared and used to identify the recombinant clones containing fungal DNA sequences not hybridizing with the DNA of the host plant, maize. Based on the nucleotide sequence information obtained from the F. moniliforme pUCF2 genomic clone, specific oligonucleotides were designed and used as primers for in vitro DNA amplification by the polymerase chain reaction. An amplification product was obtained with F. moniliforme DNA preparations whereas no amplified DNA was detected with DNAs from other fungal pathogens, including various Fusarium species, or from the host plant. This PCR analysis was successfully employed to identify F. moniliforme directly from the mycelia that develop from naturally infected maize seeds, with no need to obtain pure fungal cultures for reliable diagnosis. The protocol can be used for the diagnosis of infected plants and soils in epidemiological studies of Fusarium diseases, for seed health testing, and for evaluation of susceptibility to colonization in commercial maize hybrids.     

Suppression of the clubroot pathogen Plasmodiophora brassicae by plant growth promoting formulations in roots of two Brassica species

In this study, the ability of two organic plant growth stimulants, mainly based on algal extracts, amino acids and phosphonate, to reduce clubroot formation in the model plant Arabidopsis thaliana and two economically important Brassica species, Brassia rapa (Chinese cabbage) and Brassica napus (oilseed rape) was investigated. A commercial liquid (Frutogard^®) and a granulate (PlasmaSoil^®) formulation were used to find optimum conditions for both control of the pathogen and plant growth. Both formulations were able to significantly reduce gall formation after Plasmodiophora brassicae infection on Chinese cabbage, but PlasmaSoil^® gave better effects, possibly as a result of the continuous supply of the components to the soil. Individual components did not have the same effect. Clubroots on oilseed rape could also be reduced. In contrast, club formation was not reduced in the model plant Arabidopsis thaliana. This points to interesting differences in the induction of resistance in the different species. Cross‐sections of infected and treated roots stained for different macromolecules (callose, lignin, suberin) indicated differences in anatomy as a result of the two formulations. The results indicate an application for the granulate formulation PlasmaSoil^® in clubroot control.     

Detection of wheat-barley translocations by genomic in situ hybridization in derivatives of hybrids multiplied in vitro

Wheat-barley translocations were identified by genomicin situ hybridization (GISH) in backcross progenies originating from in vitro regenerated wheat (Triticum aestivum L. cv. Chinese Spring) × barley (Hordeum vulgare L. cv. Betzes) hybrids. The regenerated hybrids were pollinated with the wheat line Martonvásári 9 kr1. Five translocated wheat-barley chromosomes were recovered among 51 BC₂F₂ progeny from the in vitro regenerated wheat × barley hybrids. All were single breakpoint translocations with the relative positions of the breakpoints ranging from the centromere to about 0.8 of the relative arm length. Of the four translocations with intercalary breakpoints, three were transfers of terminal barley segments to wheat chromosomes; one was a transfer of a terminal wheat segment to a barley chromosome. Because of the absence of diagnostic N-bands, the identity of three barley segments could not be determined; in one translocation the barley chromosome involved had a NOR so it must have been 5H or 6H, and the centric translocation was 4HS.2BL. Following selfing, homozygotes of four translocations were selected. The experiment suggests that in vitro culture conditions are conducive for major genome rearrangements in wheat-barley hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.