Influence of regeneration and selection procedures on the production of aluminium tolerant potato regenerants

Summary Tests were made of the influence of selective and nonselective culture media on the rate of production of aluminium tolerant regenerant clones. Regenerants died when selective medium containg Al was continuously applied in tolerance tests. The production of aluminium tolerant clones was dependent on alternating selective and nonselective media. Tests using a medium containing 6 mmol al resulted in 50% growth as compared to control plants. Regenerants showed no growth when a medium with 10 mmol Al was applied. No constant tolerance was obtained when only a nonselective medium was used for regneration.     

离体筛选耐盐碱马铃薯品种试验

试管模拟盐碱土对30个马铃薯品系材料进行了耐盐碱筛选。结果表明，当培养基中总盐含量达到0．1％时，耐盐碱能力最差的品系就开始死亡，如144－4、3－2、65等品系，而耐盐碱能力强的品系，即使总盐含量达到0．4％，植株仍能存活，如品系I、131、31等。耐盐碱能力强的品系，其培养基的pH值在培养过程中都逐渐下降，到取样时均达到了适宜植株生长的范围。这说明，耐盐碱的植株能够向土壤中分泌大量H＋。     

Oryzalin Treatment of Potato Diploids Yields Tetraploid and Chimeric Plants from which Euploids could be Derived by Callus Induction

Shoot apices from six diploid potato genotypes for polyploidisation were treated with oryzalin and colchicine solutions at different concentrations and incubation times. In addition to chimeric plants, diploids, one octoploid and completely ploidy doubled plants (4x) were regenerated in these experiments. Tetraploid plants were obtained for three of the six genotypes treated with oryzalin, and no tetraploid plants were obtained after colchicine treatments of any of the genotypes. The best result of 43% tetraploid plants was achieved using 28.8 μM oryzalin and 24 h of incubation with the L37 genotype. Moreover, we determined that doubled plants should not be selected on the basis of a single measurement and require additional ploidy checks in successive clonal generations. Some L37 plants that were initially identified as tetraploids by flow cytometry produced diploid or mixoploid plants in the next clonal generation. Morphological characters of doubled L37 plants were compared with the original genotype. The PVY resistance present in a parental clone was maintained in all regenerants and significant differences among ploidy variants were found for some quantitative characters. Molecular AFLP analyses did not show any differences among genotypes. Since polyploidizing substances may produce a large proportion of chimeric plants, we applied a method to increase the number of tetraploid plants. Callus culture of chimeric leaf explants was performed on regeneration medium and tetraploid shoots were obtained in four of the six genotypes tested. The best result of 40% tetraploid plants was achieved with explants from the L37 genotype. This method was successful in producing additional tetraploid genotypes from mixoploid plants obtained after oryzalin treatments.     

Resistance to late blight (Phytophthora infestans (Mont.) de Bary) of potato plants regenerated fromin vitro selected calli

Summary Cells of seven potato cultivars were selectedin vitro with culture filtrate (CF) ofPhytophthora infestans (Mont.) de Bary. Regenerated plants were tested for late blight resistance. The aim of the study was to check the efficacy of CF in the selection of potatoes for resistance to late blight and to evaluate the effects of additive factors on general resistance. One selection cycle, applied to a cell culture system, selected cells resistant to toxic metabolites of CF.In vivo screening of clones regenerated from selected cells was done in two steps: on whole plants, assessing foliage late blight, and on detached leaves, assessing single factors of horizontal resistance. In general, the frequency of resistant variants selected with CF did not differ from that of resistant somaclonal variants. Nevertheless, high concentrations of CF in the growth medium seemed to induce an improvement of some partial resistance factors as compared with the source plants.     

电子束辐射诱变及克隆选择培育好好芭抗寒无性系的研究

利用电子辐射处理好好芭试管苗，从改变基因型入手提高其抗寒力，以获得稳定遗传的好好芭株系。研究表明150GY的辐射量是诱发好好芭发生变异的最佳剂量，结合生化标记，抗寒相关的酶类——淀粉酶和过氧化物酶活性分析，人工模拟低温实验的克隆检测，筛选出了具有较高抗寒能力的好好芭株系，这些株系通过无性系繁殖可用于抗寒育种。     

Response of potato cell cultures to aluminium and selection of tolerant cell lines

Summary We developed a method for the selection of aluminium-tolerant cell lines by using a liquid medium which more closely simulates acid soil conditions than media used previously. This medium has a pH of 4.8 and contains Al₂(SO₄)₃ instead of Al-EDTA as the selective agent to avoid the toxic effects of EDTA. It is shown that Al₂(SO₄)₃ exhibits a similar toxic effect on the growth of intact plants and cultured cells. With this medium, potato cell lines tolerating 2 mmol/l aluminium have been selected.     

Genotypic Difference of Sorghum bicolor in the Callus Formation and Callus Growth on Aluminum–Containing Medium

Abstract

We examined the genetic variation in the callus performance of the Al–tolerant and Al–susceptible genotypes of sorghum (Sorghum bicolor L.) on Al–containing medium. Addition of Al to the medium with a low pH, low Ca and low phosphate resulted in a severe decrease in the percentage of callus formation (GF) in both Al–tolerant and Al–susceptible genotypes. However, the percentage of callus formation in the expiants from Al–tolerant genotypes was higher than that from susceptible genotypes on Al–containing medium. The genotypic difference in the relative growth of calli on Al–containing medium and in the suppression of callus formation by Al was in agreement with that in the hematoxylin staining score. Al might have been considered as a limiting factor that affected the callus growth. There was a significant correlation (r= –0.664*) between the percent relative suppression of callus formation by Al and the percentage of callus formation on Al–containing medium. The callus growth in fresh weight and size on Al –containing medium relative to that on Al–free medium (relative growth on Al–containing medium) was greater in Al–tolerant genotypes than in the other genotypes. The relative growth of callus in fresh weight and size on Al–containing medium significantly correlated (r=–0.820** and r=– 0.956**, respectively) with the percent relative suppression of callus formation by Al. The tissue culture is considered to be a useful tool for selecting Al–tolerant haploids at the callus stage.     

农杆菌介导法将抗草甘膦基因转入玉米自交系

利用农杆菌介导法将抗草甘膦基因(ssu-G₂-aroA)转入优良玉米自交系R18-599和齐319,侵染后的愈伤组织转至潮霉素浓度为10 mg/L的筛选培养基上,继代筛选3轮,每轮3周;得到的抗性愈伤组织在含有潮霉素浓度为3 mg/L的分化培养基上进行分化,R18-599获得30株再生植株,齐319获得18株再生植株。经PCR鉴定后共得到7株转化抗性植株(4株R18-599、3株齐319)。经PCR-Southern及RT-PCR检测结果表明,ssu-G₂-aroA基因已稳定整合到了玉米基因组中,并在RNA水平上得到表达。     

小麦-中间偃麦草蓝粒代换系的创制与鉴定

小麦的蓝粒性状可作为表型标记用于小麦育种和遗传学研究,来自中间偃麦草的蓝粒种质材料尚鲜见报道。本研究通过八倍体小偃麦中5 (2n=8x=56, AABBDDXX)与中国春缺-四体系列材料杂交,在中5×N4BT4A和中5×N7BT7D杂交组合后代中获得了两份蓝粒材料,编号分别为Zh5-a2-1和Zh5-c13-2。利用细胞遗传学和分子标记方法对这两份蓝粒材料进行了染色体组成分析。以中间偃麦草基因组DNA为探针的GISH分析显示,这两份蓝粒材料的染色体数均为2n=42,包括40条小麦染色体和两条中间偃麦草染色体。利用重复序列探针pSc119.2和pAs1进行的FISH分析表明,Zh5-a2-1和Zh5-c13-2均为二体代换系,被代换的一对小麦染色体分别为4B和4D。通过用St、E~e和E~b基因组DNA作探针进行GISH分析,证明这两份蓝粒代换系中的中间偃麦草染色体均为St组染色体,但与中5中的中间偃麦草染色体比较发现这对St组染色体的短臂端部发生了缺失。利用二倍体长穗偃麦草E~e基因组的SNP标记分析证明,两份蓝粒代换系中的中间偃麦草染色体与长穗偃麦草的4E~e染色体同源,即Zh5-a2-1和Zh5-c13-2分别为4St(4B)和4St(4D)代换系,命名为SubZh5-4St(4B)和SubZh5-4St(4D)。同时说明,中间偃麦草的4St染色体上带有蓝粒基因。通过对450个小麦SSR标记进行筛选,获得了4个可跟踪鉴定4St染色体的特异SSR标记。研究结果可用于蓝粒小麦品种的培育和中间偃麦草蓝粒基因的遗传学研究。     

Resistance toErwinia carotovora subsp.atroseptica in second backcross populations of somatic hybrids betweenSolanum tuberosum andS. brevidens

Summary Eleven clones obtained from a cross between cv. Katahdin and fusion products betweenSolanum tuberosum and the non tuber-bearing speciesS. brevidens, were backcrossed again withS. tuberosum (clone AR80-127-5). Small tubers harvested from 583 seedlings of these second backcross (BC2) populations were screened for tuber tissue resistance toErwinia carotovora subsp.atroseptica under aerobic conditions. After multiplication in the field. BC2 clones again were screened for soft rot resistance under aerobic as well as anaerobic conditions. In general, the resistance of the BC2 populations was reduced in comparison with the BC1. Variation for resistance was found within the BC2 populations, but there was no correlation between the results of the three tests performed. Four BC2 populations were also screenned for resistance to blackleg in the field. Significant differences were found between populations for mean percentage of diseased plants, but these differences could not be explained by the resistance of the parental clones.     

Evaluation in pot and field trials of resistance of potato cultivars and breeding lines to common scab caused byStreptomyces scabies

Summary The resistance to common scab of nine cultivars and eight advanced breeding lines of potatoes, fiveSolanum andigenum Juz. et Buk. clones, and three potato breeding lines selected for resistance to common scab, was investigated in a field and a pot trial. In the pot trial potaroes were planted in inoculated coarse river sand in plastic pots (150 mm in diameter) placed on irrigated subsoil in a glassfibre house. In a field trial potatoes were planted in an artificially infested field. The harvested tubers were scored individually for scab incidence according to two scales, viz. a 1–5 point scale for percentage surface covered with scab and a 1–3 point scale for lesion type. A scab index value was calculated for each tuber. A significant correlation coefficient of 0.82 was obtained for the results of the two tests.     

Performance and stability of nine 4x clones from 4x−2x crosses and four commercial cultivars

Summary Nine clones from 4x−2x crosses were evaluated along with four cultivars in six Wisconsin environments. The 4x clones from 4x−2x crosses had as their 2x parents hybrids with haploid Tuberosum and either Group Phureja orSolanum tarijense. The 2x hybrids produced 2n pollen by first division restitution. The traits evaluated were marketable yield, mass density and chip color. Stability analysis was conducted by determining the linear relationships between mean yield of individual clones and environmental mean yields. Mean square deviations from linear regression (s² _d) were used to evaluate phenotypic stability and regression coefficients (b) as environmental response parameters. Three 4x clones from 4x-2x crosses were more stable and not significantly different for yield than cv. Atlantic; some also had superior specific gravity and chip color. The results suggest that the 4x×2x breeding method is very efficient for the production of high yielding, stable clones with superior processing qualities, particularly since the 4x clones from 4x-2x crosses originated from very small source populations.     

Male sterility and 2n pollen in 4x progenies derived from 4x×2x and 4x×4x crosses in potatoes

Summary The objectives of these experiments were to evaluate male sterility (MS) and 2n pollen frequency in 77 4x×2x families and 26 4x×4x families. The 2x parents were haploid-species hybrids and the 4x parents mainly clones withS. tuberosum ssp.tuberosum (tbr) andS. tuberosum ssp.andigena (adg) in their genetic background. The female parents had different cytoplasms: adg, tbr,S. demissum (dms), andS. stoloniferum (sto). Families from female parents with adg or dms cytoplasm did not have MS plants. However, families derived from crosses of many tbr females with 2x hybrids and a 4x adg clone had a high percentage of MS plants; all the progenies from cv. Serrana (sto cytoplasm) possessed tetrad sterility. These results can be explained as due to an interaction between a dominant male sterility (Ms) gene with tbr and sto cytoplasms. Some MS 4x hybrids (tbr×adg) used as females had the Ms gene, but male fertility was restored in some plants of their progenies when they were crossed with 4x tbr clones. This indicates that some tbr clones have a fertility restorer gene (Rt). The results from both 4x×2x and 4x×4x families fit the expected ratios assuming chromatid segregation for both the Ms and Rt loci. The gene frequency of parallel spindles was estimated in the 4x population as 0.74.     

红粒春小麦穗发芽抗性鉴定及相关分子标记的有效性验证

为了筛选出能鉴定红粒小麦穗发芽抗性的分子标记及抗穗发芽的种质材料,检测了67份红粒春小麦品种(系)的发芽指数,并利用4个与穗发芽抗性相关的标记(Vp1B3、Xgwm155、Xwmc468和Xgwm397)对这67份品种(系)进行了PCR扩增,分析了扩增片段与发芽指数的关系.结果表明,在67个红粒春小麦品种(系)中,12...     

Growth analyses of eleven potato cultivars grown in the greenhouse under long photoperiods with and without heat stress

Six potato cultivars from North America were grown in two greenhouses under 18-h photoperiods. All plants were started in a greenhouse that was well ventilated and maintained as close as possible to summer ambient temperatures. One month after planting, half the plants of each cultivar were moved to a greenhouse that was kept at 30 C during the night and was allowed to reach 40 C or higher during the day. Plants from each treatment were harvested at monthly intervals for growth analyses. Concurrently, five clones considered to be heat tolerant were subjected to a similar test in a second experiment. In response to high temperatures, plants in both experiments were taller with smaller leaves, lower tuber yields, higher ratios of stem to leaf dry matter, and lower ratios of tuber to leaf and to stem dry matter. Tubers were lower in percent dry matter, maturity was delayed, and cuttings developed fewer tubers. Cultivars differed greatly in the degree to which they were affected by the heat. In the first experiment, Norchip had outstanding heat tolerance. LT-2 was the most tolerant of the clones in the second experiment. For the cooler treatment the best performance in either experiment was from two clones originally selected in the lowland tropics.     

An evaluation of the effect of Bacillus cells and Bacillus spores in association with cowpea granules as starter cultures for the fermentation of African oil bean (Pentaclethra macrophylla Bentham) to `ugba'

Studies on the improvement of the traditional production of `ugba',a protein-rich fermented African oil bean seed product, were undertaken,by developing starter cultures of Bacillus subtilis cells andspores in association with cowpea granules. The viability of the cellsin association remained stable at 94.5% for 6 months at 30 <sup>°</sup>C and for up to 10 months at 4 <sup>°</sup>C while the viability of the sporesin association remained stable at ca. 96% for up to 10 months at both4 and 30 <sup>°</sup>C. The starter cultures resulted in high increases inprotease activity from ca 2.8 mg N/min to about 51.6 ± 0.4 mg N/minin 48 h and a corresponding increase in amino-nitrogen content of ca2.0 ± 0.2 mg N 100 g dry matter (DM) to ca 18.5 ± 0.3 mg N/100g (DM) during the same period. Changes in the protease activity of thenatural process were gradual and increased from 3.0 mg N/min to 38.0 ±0.8 mg N/min after 5 days of fermentation. The maximum amino nitrogencontent of `ugba' produced by the starter cultures (18.5 ± 0.3 mgN/100 g DM) after 2 days was significantly (p < 0.05) higher than themaximum amino nitrogen content (12.5 ± 0.8 mg N/100 g DM), of `ugba'obtained by the natural process. `Ugba' produced by the starter cultureswere well accepted and compared favorably with the natural product.     

北方冬麦区与黄淮北片优良小麦品种(系)耐热性评价

为筛选小麦耐热种质资源,在开花后15d开始覆盖塑料大棚,模拟高温胁迫,对来自我国北方冬麦区和黄淮北片主推的53个小麦品种(系)的千粒重热感指数、容重热感指数、几何平均产量进行分析,并以耐热品种TAM107为对照,结合苗期细胞膜热稳定性对参试材料的耐热性进行评价。结果表明,在三年(2008-2011)试验中,千粒重热感指数均小于1的品种(系)有9个(05CA306、京冬8号、邯6228、衡6632、农大3492、农大3432、济麦19、农大212和衡4399),容重热感指数均小于1的品种(系)有8个(衡观216、农大3492、农大413、京冬8号、山农2149、农大212、衡6632和济麦19)。依据参试品种(系)在3年的几何平均产量及热感指数的结果,农大212、农大3492、京冬8号、济麦19、衡6632、农大413、山农2149和衡观216可作为耐热高产品种(系)在生产上推广应用。利用细胞膜热稳定性筛选出44个苗期耐热性好的品种(系)。综合细胞膜热稳定性法及田间评价结果,农大212、衡6632、农大3492、济麦19、农大413、山农2149和衡观216为耐热性较好的品种(系)。     

一株耐铝甜瓜枯萎病拮抗细菌的分离鉴定及其耐铝特性研究

从甜瓜根际酸性土壤中分离筛选到1株耐铝甜瓜枯萎病拮抗菌，命名为A2。根据表型、生理生化特征和16S rRNA基因序列相似性分析，将其鉴定为Pseudomonas sp.。菌株A2对甜瓜枯萎病病原菌的相对防效为68.3%，且拮抗能力具有遗传稳定性。相比AlCl3处理，菌株A2对Al2(SO4)3表现出了更好的耐受性，最高可耐受50mmol/L Al3+。在含有A13+的S-LB培养基中，菌株最适生长温度为30 ℃；培养基初始pH值的降低会加剧A13+对菌株A2的毒害作用。研究结果可为含活性铝的酸性土壤中甜瓜枯萎病的生物防治提供优良菌株资源和理论基础。     

Breaking primary dormancy in seeds of the perennial pasture legume tedera (Bituminaria bituminosa C.H. Stirt. vars albomarginata and crassiuscula)

Tedera (Bituminaria bituminosa vars. albomarginata and crassiuscula) is a perennial pasture species with agronomic characters ideally suited to Mediterranean climates. Tedera seed has a period of after-ripening or primary dormancy typically lasting three months, which delays assessment and breeding of elite hybrid varieties. Temperature, chemical and mechanical methods were investigated in conjunction with in vitro culture to circumvent this dormancy period across a range of parental and hybrid genotypes. Temperature treatment of T5 (Tedera accession 5) and T48 (Tedera accession 48) alone was not sufficient to break dormancy (24.0% and 14.7% germination); however, when combined with soaking in gibberellic acid (GA₃) and mechanical scarification resulted in 79.7% and 84.3% germination respectively. In an effort to further improve this result for valuable hybrid genotypes, we combined mechanical scarification with removal of seed coat after imbibition and in vitro culture on B5 medium until radicle emergence. This resulted in breaking dormancy from 96% to 100% of parent seeds and 100% of hybrid seeds. Hardening the germinated F₁ or F₂ seedlings 4 d after first transfer to in vitro culture resulted in 100% survival of plants to soil. This procedure is now used on a routine basis in the Australian tedera breeding programme.     

杜鹃红山茶离体快速繁殖

以杜鹃红山茶实生小苗为试验材料,探讨离体快速繁殖的方法。结果表明:茎段外植体在MS+BA 1 mg/L+IBA 0.01 mg/L+GA3 10 mg/L的培养基上培养60 d,侧芽萌发率高达68%;将这些萌发的侧芽连同原来的茎段外植体移至添加BA 0.2～0.4 mg/L的培养基培养1个月,74%的侧芽能够继续生长并形成丛生芽;再将丛生芽移至1/2 MS+IBA 4 mg/L+0.1%活性炭的培养基上培养1个月,得到大量伸长的芽条;将芽条切离母体后插植于1/2 MS+IBA 8 mg/L的培养基,60 d后生根率达到90%。通过以上过程繁殖得到的小苗质量良好,移栽1个月后的成活率可达90%。