Enhancement of RAPD Analysis by Restriction-endonuclease Digestion of Template DNA in Wheat

Random amplified polymorphic DNA (RAPD) analysis has proven to be an effective procedure for molecular marker applications in plant breeding, although non-specific amplification may limit its utility in some species. The objective of this study was to determine the effectiveness of restriction-endonuclease digestion of template DNA for elimination of non-specific amplification and generation of heritable RAPD markers. Restriction endonucleases digested wheat DNA to completion in amplification buffer, suggesting that the restriction endonuclease can be added directly to the reaction mixture prior to amplification. A 1-h 37°C step was programmed into the thermal cycler for restriction-endonuclease digestion which was followed immediately by amplification. Non-specific amplification was reduced and DNA marker patterns were altered in digested samples when compared to undigested samples. Genetic segregation of two polymorphic markers tested in F₅ inbred progeny fit expected 1:1 ratios. These results suggest that heritable DNA markers may be generated with reduction in non-specific amplification when restriction-endonuclease digestion of template DNA is conducted as part of the RAPD procedure.     

In vitro androgenesis of wheat: from fundamentals to practical application

Wheat anther cultures have a history of almost 30 years and are nowemployed efficiently in many countries of the world for the developmentof doubled haploid lines for breeding. The present paper discusses keyquestions of the elaboration and perfection of the method: cytologicalaspects of in vitro androgenesis, the conditions required for theembryogenic development of microspores and the applicability of anthercultures in the Martonvásár wheat breeding programme.     

Metaphase-I analysis of a Triticum aestivum x T. monococcum hybrid by the C-banding technique

Summary The meiotic pairing behaviour at metaphase I of a Triticum aestivum×Triticum monococcum hybrid has been studied by means of the C-banding technique to ascertain the homology between the chromosomes in the A genome of the two species. The technique allowed the A and B genome chromosomes and the 2D, 3D and 5D chromosomes to be identified. Differences in the level of chromosome pairing in the A genome were noted. The T. monococcum 4A chromosome did not pair with any of the T. aestivum chromosomes in any of the metaphase I cells analysed. Two reciprocal translocations between the 2B and 2D chromosomes on one side and the 2A and 3D on the other side have been identified. The usefulness of the C-banding technique in the study of chromosome homology among species related to wheat is discussed.     

Meiotic studies of spontaneous hybrids of Amaranthus: genome analysis

The meiotic behaviour of 13 spontaneous interspecific F₁ hybrids of Amaranthus was studied. The hybrids between species with n= 16 chromosomes had 16 bivalents but varied considerably in pollen stainability (0–55%). These results suggest the existence of cryptic structural hybridity. The hybrids involving A. cruentus (n = 17) and species with n = 16 (A. caudatus and A. quitensis) always formed 15II+1 III with very low pollen stainability (5–7%). Further observations indicated that Amaranthus species are allotetraploids with basic numbers of x= 8 and x= 9 but exhibit x= 16 and x= 17 as secondary basic numbers, as demonstrated by (a) the frequent presence of 811 + 171 in the meiosis of the hybrid A. spinosus (n = 17) × A. hybridus (n= 16); and the occurrence of secondary associations between bivalents in MI. Genomic formulae are proposed for each species, on the basis of the meiotic behaviour of the hybrids studied.     

花生基因组SRAP-PCR体系的优化

【研究目的】研究花生基因组SRAP-PCR的扩增条件,建立其优化扩增体系;【方法】对模板DNA、引物、dNTP mixture、Taq DNA聚合酶浓度及退火温度设置不同梯度,研究各因素对PCR结果的影响;【结果】扩增程序为:94℃变性2min,1Cycle;94℃变性30s,48℃复性30s,72℃延伸1min,40Cycles;72℃延伸7min。反应体系成份为:DNA模板80ng,左右引物各200ng,dNTP mixture2.0mmol/L,Taq DNA聚合酶3U,10×Buffer8μl,总体积60μl。【结论】建立了满足花生基因组SRAP-PCR的优化扩增体系。     

Marker‐assisted backcross breeding to combine multiple rust resistance in wheat

A widely grown but rust susceptible Indian wheat variety HD2932 was improved for multiple rust resistance by marker‐assisted transfer of genes Lr19, Sr26 and Yr10. Foreground and background selection processes were practised to transfer targeted genes with the recovery of the genome of HD2932. The near‐isogenic lines (NILs) of HD2932 carrying Lr19, Sr26 and Yr10 were individually produced from two backcrosses with recurrent parent HD2932. Marker‐assisted background selection of NILs with 94.38–98.46% of the HD2932 genome facilitated rapid recovery of NILs carrying Lr19, Sr26 and Yr10. In the BC₂F₂ generation, NILs were intercrossed and two gene combinations of Lr19+Yr10, Sr26 + Yr10 and Lr19+Sr26 were produced. A total of 16 progeny of two gene combinations of homozygous NILs of HD2932 have been produced, which are under seed increase for facilitating the replacement of the susceptible HD2932 with three of the sixteen improved backcross lines with resistance to multiple rusts.     

Comparison of nuclear DNA content of citrus rootstock populations by flow cytometry analysis

Citrus species are widely grown in the world. Plant characteristics of the rootstock populations used for orange, lemon, mandarin and grapefruit cultivars should be known as well as those of the cultivars that help true‐to‐type nursery plant production. In this study, cell nuclei were isolated from leaf tissues of seedlings of trifoliate orange, sour orange, rough lemon,‘Volkamer’ lemon,‘Cleopatra’ mandarin,‘Hyokan’, ‘Sanbokan’, ‘Kinkoje’, ‘Carrizo’ citrange and ‘Swingle’ citrumelo, and then fluorescence intensities were measured on propidium iodide‐stained nuclei by flow cytometry. Nuclei isolated from the triploid ‘Tahiti’ lime with a known nuclear genome size were used as the internal standard to estimate the nuclear DNA content of Citrus seedling populations in absolute units. Results obtained from cytograms and histograms indicated that all seedlings analyzed were diploid. In addition, differences between the species for nuclear DNA content were also found to be significant. ‘Hyokan’ seedlings had the biggest genome size, 0.984 pg/2C, whereas trifoliate orange seedlings had the smallest genome size, 0.678 pg/2C. Flow cytometry analysis could be used for obtaining accurate and rapid results for cytological observations of seedling populations of Citrus.     

Intergeneric hybrids and an amphiploid between Elymus canadensis and Psathyrostachys juncea

Summary Fifty four hybrid plants between Elymus canadensis and Psathyrostachys juncea were obtained by handpollination and embryo culture. The average cross compatibility between both species was 31.2 percent. One amphiploid plant was induced by colchicine treatment. The hybrid and amphiploid plants resembled P. juncea in appearance but showed a higher plant height and dry matter yield than the parents. The hybrids showed extremely low pollen stainability and were completely sterile. With the exception of one plant (2n=3x+1=22), all hybrid plants were allotriploids (SHN, 2n=3x=21). The amphiploid plant (SSHHNN, 2n=6x=42) showed 58.9% pollen stainability and 11.6% seed fertility.Mean chromosome associations of the hybrids and amphiploid at metaphase I were 0.02IV+0.06III+2.03II+16.91I and 0.07III+18.00II+5.85I, respectively. Lagging chromosomes, chromosome bridges, abnormal cytokinesis, and micronuclei were occasionally observed at the anaphase, telophase, or tetrad stage.     

Genomic Relationships of Triticum sharonense with Other S-Genome Diploid Triticum Species

Triticum sharonense was hybridized with autotetra-ploid T. speltoides, T. longissimum and T. bicorne. Meiotic analysis of these hybrids showed that T. sharonense is almost equally related to both T. speltoidcs and T. longissimum, while it is comparatively distant from T. bicorne. Therefore, this study does not support treating T. sharonense as a subspeies or variety of T. lottgissimum.     

Characterization of the durable resistance to yellow rust in old winter wheat cultivars in the Netherlands

Summary Winter wheat cultivars released in the Netherlands before 1930 carried durable resistance to yellow rust. Cultivars released in the period between 1930 and 1950 often were durably resistant while recent cultivars infrequently showed durable resistance. This durable resistance was not difficult to transfer to new cultivars. Twenty nine older cultivars with durable resistance and eight recent non-durably resistant cultivars were tested in the seedling stage and in the adult plant stage against 12 West-European yellow rust races and against some non-European races in the seedling stage only. The adult plant tests were carried out in race nursery tests in the Flevopolder. Per race nursery all 37 cultivars, planted in hills of about 20 plants on both sides of the highly susceptible cv. Michigan Amber, were exposed to one race.The infection type of each cultivar-race combination was scored on 0 to 9 scale once in the seedling stage and twice in the adult plant stage. In the race nurseries the percentage leaf area affected was evaluated three times to be used to calculate the area under the disease progress curve (AUDPC). This AUDPC multiplied with the mean infection type in the field gave the susceptibility index (SI).The infection types were classified into resistant (R), intermediate (I) or susceptible (S) when the infection types were 0 to 3, 4 to 6 or 7 to 9, respectively. Four categories of resistance were discerned on the basis of the three infection type scores: 1) RRR, overall resistance; complete or near-complete resistant at all stages. 2) SRR, adult plant resistance, complete- or near-complete resistant at the adult plant stage only. 3) SRS and SSR, temperature sensitive resistance, the resistance changed from the one evaluation data to the other. 4) SSS and an SI lower than that of Michigan Amber, partial resistance.The frequencies of overall, adult plant and temperature sensitive resistance were 1.4, 52 and 54% in the older cultivars and 40, 62 and 22% in the recent ones, respectively. Among the older cultivars all had a fair to high level of partial resistance, the SI being on average only 20% of that of Michigan Amber, while most cultivars also seemed to carry temperature sensitive resistance. The partial resistance of the recent cultivars was of a much lower level with a mean SI compared to that of Michigan Amber of 61%. Partial resistance was highly correlated (r = –0.94) with the mean resistance scores from the Dutch Recommended Cultivars Lists. It was concluded that partial resistance and temperature sensitive resistance were the major components of the durable resistance in the older cultivars.