The potential of selection and breeding for improved salt tolerance in lucerne (Medicago sativa L.)

Summary Four cultivars of Medicago sativa L. were subjected to selection for improved salt tolerance using a salinized solution culture technique at 250 mM NaCl. Selections were made after two weeks growth, based upon seedling shoot length differences. High (shoot lengths 14–31 mm) and low (7–12 mm) selection lines were established. Unselected shoot lengths ranged from 0–11 mm. Selection intensities ranged from 0.17% to 0.22% for the high selection lines, and from 0.30% to 1.05% for the low selection lines.Eleven selected plants from cv. CUF 101, and 10 from Local Syria were grown on and polycrossed. A sample of progeny seed from each was grown at 8 NaCl concentrations in solution cultures. Selected line seedlings produced shoots at 225, 250, 260, and 275 mM NaCl, whereas unselected material failed to grow above 225 mM NaCl. A second cycle of selection at 280 mM NaCl with these two cultivars achieved selection intensities of 0.21% and 0.45% for high and low selection lines, respectively. Selected lines grew and produced shoots at 300 mM NaCl.Seedlings derived from 1st and 2nd selection cycles and unselected control material were grown for 6 weeks at 0, 150, 175, and 200 mM NaCl in a sand culture experiment. The high selection line produced significantly greater shoot fresh weight, dry weight, and % live shoot weight than the unselected control material. Clearly selection as practised here can isolate individual seedlings having enhanced genetically based tolerance to NaCl, which is manifest in mature plants derived from polycrossing those selected individuals.     

The application of in vitro tiller induction in Lolium multiflorum

Summary Eight Lolium multiflorum genotypes were micropropagated for over one year. Using Murashige & Skoog's basal medium, various growth regulators (6-benzylaminopurine, indole acetic acid, adenine sulphate) were tested for their effect on micropropagation. Plantlet handling methods, culture conditions and culture procedures were also compared. Differences were observed in rates of micropropagation between plant genotypes. Under the best conditions for micropropagation, an average of approximately 50 tillers were produced in a 5–6 week period, compared with an estimated 8–15 tillers from soil-grown plants over the same period. Tillers must reach a certain developmental stage before they will grow independently, therefore the number of surviving propagules at subculture under micropropagation and in soil is about half the number of tillers produced. A proposed system for the micropropagation of Lolium multiflorum is described.     

Plant regeneration from callus culture in potato

Summary A procedure for plant regeneration from callus culture of potato, Solanum tuberosum L. is described. Calli were induced from 1–2 mm long shoot apices of potato cultivars Cara and A25/19 on half-strength Murashige and Skoog's medium (half-MS) supplemented with 3.2 mg IAA (indole-3-acetic acid), 1.0 mg kinetin (6-furfurylamino)purine], and 0.5 mg 2,4-D [2,4-dichlorophenoxy)acetic acid]/1. Sixty percent explants produced nodular calli on this medium within 30 days. Calli differentiated into shoot-primordia when subcultured on half-MS medium supplemented with 0.5 mg 2,4-D and 1.0 mg zeatin [6-(4-hydroxy-3-methybut-2 enylamino)amino purine]/1. Differentiated calli on half-MS medium without growth hormones produced complete plantlets which were cloned on the same medium and transferred into soil.     

Induction of mutations for heat tolerance in potato by using in vitro culture and radiation

Heat tolerant mutants were obtained in two commercial potato cultivars, `Kufri Jyoti' and `Kufri Chandramukhi' through in vitro mutagenesis of in vitro propagated plantlets. Gamma-irradiated (20 and 40 Gy) shoots were micropropagated for three cycles (M₁V₃). A large number of the micropropagated shoots produced microtubers at 28 ^°C. Microtubers induced at high temperature had distorted shape but showed normal germination in field. Under stress conditions of high temperature, the frequency of chlorophyll variants increased in the gamma irradiation-derived material, however, nearly 40% of the plants had normal leaf tissue, whereas control plants showed completely damaged leaves. This revised version was published online in July 2006 with corrections to the Cover Date.     

Crown depth and susceptibility to crown rot in wheat

Sources of partial resistance to crown rot caused by Fusarium pseudograminearum are detected in mature plants grown in artificially inoculated soil in the field. The resistance in most but not all of these sources can also be detected in seedlings. In order to determine whether partial resistance is related to depth of crown formation, this character was measured in 13 cultivars/lines with a range of reaction to crown rot. It was also measured in doubled haploid plants from the cross, Batavia/2–49. Crown depths varied from 17.1 mm to -2.3 mm (above ground) in pots in a waterbath at 25 ^°C and from 43.5 mm to 20.7 mm when plants were grown in the field. The correlation coefficient between relative susceptibility to crown rot (Field test) and crown depth of 13 cultivars/lines was 0.57 (p ≤ 0.05). With the exception of the cultivars, Sunco and Pelsart, partial resistance to crown rot was inversely related to depth of crown formation. This indicates that depth of crown formation may be partly responsible for the reaction of a cultivar/line to crown rot. Depth of crown formation was also measured in susceptible and partially resistant cultivars/lines grown from seed planted at different depths. As depth of seeding increased, depth of crown formation in partially resistant and susceptible cultivars/lines increased in similar proportions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Potential of a Fusarium eumartii culture filtrate on the screening for wilting resistance in potato

Summary Fusarium solani f.sp. eumartii Carp. Snyder and Hansen (Fusarium eumartii) is a soil inhabitant that induces the so-called Potato Wilt and Stem End Rot disease. Prior to wilting, the pathogen induces peculiar small bronze spots on the leaflets. Failure to isolate F. eumartii from infected leaflets suggests the involvement of a toxin in the disease. The fungus was grown in liquid Richard's medium and thereafter a filtrate was obtained dialyzing (MW cutoff 12,000–14,000) and sterilizing the culture by filtration (0.22 m). Potato leaves treated with both the pathogen or the filtrate showed symptoms of bronze spots and significantly higher electrolyte leakage when compared to controls. Tomato leaves showed neither bronze spots nor electrolyte leakage after plant inoculation with the pathogen or with the filtrate treatment. Both, the absence of visible symptoms and the lack of electrolyte leakage in tomato could be associated to a certain degree of host specificity of the F. eumartii filtrate towards potato. The filtrate also induced symptoms similar to infections by F. eumartii in adult plants and in vitro plantlets of cultivars Huinkul MAG and Kennebec. Callus responses to the filtrate were related to responses of the cultivars to the pathogen in greenhouse. These results show the potential of the culture filtrate of F. eumartii for use in screening for wilting resistance.     

Evaluation of the U.S. cucumber germplasm collection for root size using a subjective rating technique

Summary Root size in cucumber (Cucumis sativus L.) has received little research attention until recently. However, a large root system could improve nutrient and water uptake, and ultimately yield. Knowledge of root size could help cucumber breeders develop cultivars with improved performance. A study was conducted in the greenhouse to evaluate 857 cultigens (827 plant introduction accessions, 22 breeding lines, and 8 cultivars) of cucumber for root length. Complete data were obtained for 794 cultigens. Plants were grown from seeds in 2 replications in the greenhouse. Plants were rated for root length using a 1 to 9 scale (1–3 = small, 4–6 = medium 7–9 = large). Also, root length was measured from soil surface to the tip of the longest main root. In addition, all cultigens were evaluated for root length and dry weight to provide meaning to the subjective rating. Root rating was highly correlated with both root length (r=0.92) and dry weight (r=0.64) of roots. Root length averaged 74 mm and most cultigens (579, or 68%) had roots 40 to 110 mm long. The cultigen with the longest roots was PI 183056 (193 mm), and the shortest was PI 176954 (2 mm). The subjective rating technique provided a simple, accurate and rapid method to estimate root growth in the greenhouse.     

Identification of somatic hybrids of dihaploid Solanum tuberosum lines and S. brevidens by species specific RAPD patterns and assessment of disease resistance of the hybrids

Summary Symmetric somatic hybrids were produced by electrofusion of protoplasts of two dihaploid tuber-bearing potato (Solanum tuberosum L.) lines and Solanum brevidens Phil., a diploid non-tuber-bearing wild potato species. A total of 985 plants was obtained. Verification of nuclear hybridity of putative hybrids was based on additive RAPD patterns, general morphological characteristics and chromosome counts. 53 (90%) calli regenerated into plants which were identified as somatic hybrids. Most of the hybrids were aneuploids at the tetraploid (4×) or hexaploid (6×) level. The 20 hybrids tested expressed a high level of resistance to potato virus Y (PVY ^N ) characteristic of the S. brevidens parent. Resistance to late blight (Phytophthora infestans (Mont.) de Bary) varied between hybrids, but was on average better than that of the fusion parents. Resistance of hybrids to bacterial stem rot (Erwinia carotovora subsp. atroseptica (van Hall) Dye) was not superior to that of commercial potato cultivars.     

Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato

Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.     

Influence of genotype and culture conditions on the production of embryos from anthers of tetraploid wheat (Triticum turgidum)

Summary Anther culture of 10 tetraploid wheat (Triticum turgidum) genotypes and two backcross lines representing a wide range of genetic variation was studied in a randomized block design with three replications. Each replication consisted of 2 pots with 3 plants. The day length was 16h and temperature 25° C/15°C for day/night in a controlled greenhouse where the anther donor plants were grown. Two different treatments were used for anther culture. The first one was potato 2 medium (Chuang et al., 1978) modified by adding 0.5 mg/l glutamine and solidified by gelrite (4g/l) (Henry & De Buyser, 1981). Cultures were incubated in light (15 E m^–2 S^–1) at 26°C at 16h day length. The second medium was described by Fadel & Wenzel (1990), differing from the first by the nature of the sugar (maltose) and consistency of the medium (semiliquid by ficoll). Anther cultures were incubated in the dark at 28°C. The study of about 1300 anthers per genotype and treatment showed that both genotype and treatment affected embryo formation of tetraploid wheat. The backcross lines exhibited significant differences for androgenic abilities when compared to their common parent. Most of the genotypes were medium dependent for androgenesis and revealed significant interactions with the two treatments. Five green plantlets were regenerated and fertile doubled haploid plants were obtained from three out of the 12 studied genotypes.     

Evaluation of seedling selection for salinity tolerance in potato (Solanum tuberosum L.)

Summary Salt-tolerance in potato (Solanum tuberosum L.) was selected on the basis of germination and survival of seed sown in trays of perlite suspended in either 75 or 150 mM NaCl. Salinity reduced the germination of seed. Genetic differences in salt-tolerance were apparent with salt reducing germination more in seed collected from cv. Cara than in that collected from cv. Maris Piper. Progeny from the seedling selection were then grown to maturity to produce tubers.The relative tolerance of the parental cultivars and of unselected and selected progeny to long-term exposure to salinity was examined in a pot experiment in which plants were irrigated with either fresh water or 50 mM NaCl solution from one week after plant emergence. In this experiment, salinity significantly reduced leaf conductance, total dry matter production and partitioning of assimilate to tubers. Salinity reduced dry matter production and assimilate partitioning to tubers to a greater extent in Cara than in M. Piper. Progeny selected for short-term salt-tolerance did not exhibit greater long-term salt-tolerance than unselected progeny, and both were more sensitive than M. Piper.These results demonstrate genetic variation in salt-tolerance in potato. However, although there was a correlation between the performance of the parent to long-term salinity and survival of progeny in the seedling selection, there was no correlation between short- and long-term salt-tolerance. This suggests that characters underlying short-term tolerance may contribute to long-term tolerance but do not of themselves confer long-term tolerance. Future progress in selecting for improved salt-tolerance depends on understanding the effects of salinity on the physiological processes underlying growth and carbon partitioning.     

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.     

Influence of suboptimal temperature on biomass production of potato populations of Andean and European origin

Summary Plants grown from seed derived by crossing conventional European S. tuberosum material were compared with plants grown from seed derived by crossing S. tuberosum with various, Andean frost resistant tuberbearing Solanum species. Biomass growth at optimal (20°/10°) and suboptimal (10°/4°) temperatures was studied.Differences in increase of fresh and dry matter were found between populations of Andean and European orgins at 10°/4°. At 20°/10° no such differences were found.At suboptimal temperature, Andean hybrids produced significantly more fresh and dry matter than European crosses in two harvests (64 and 178 days old plants). Statistically, Andean hybrids were found to produce the same amount of dry matter per day at both temperatures, over the complete growth period.Height increase and flower development were strongly depressed in European crosses under the suboptimal temperatures, but much less so in Andean crosses.The Andean material appears to be an under utilized resource in potato breeding for cool climates.     

Production of intergeneric hybrid plants between <Emphasis Type="Italic">Sandersonia aurantiaca</Emphasis> and <Emphasis Type="Italic">Gloriosa rothschildiana</Emphasis> via ovule culture (Colchicaceae)

Intergeneric hybrid plants between Colchicaceous ornamental plants, Sandersonia aurantiaca and Gloriosa rothschildiana, have successfully been produced via ovule culture. After 5 days of reciprocal cross-pollination, a few pollen tubes were observed in the ovary. Although seeds were obtained in both reciprocal cross-combinations, they did not germinate under ex vitro conditions. Ovules with placental tissues isolated 14 days after cross-pollination of S. aurantiaca × G. rothschildiana were cultured on a medium containing 0.01 mg l^–1 each of -naphthaleneacetic acid (NAA) and 6-benzyladenine (BA), on which 41.5% of ovules swollen and produced callus-like structures within 10 weeks. When such swollen ovules were transferred to a medium containing 0.1 mg l^–1 each of NAA and BA, 7.5% of the initially cultured ovules produced rhizome-like structures within 6 weeks. Among the rhizome-like structures, those derived from two independent ovules (3.7% of the initially cultured ovules) produced multiple shoots following transfer to a medium containing 0.25 mg l^–1 NAA and 2.5 mg l^–1 BA. Multiple shoot-derived plantlets were established on a plant growth regulator-free medium, and they were successfully transplanted to pots. Early verification of their hybridity was accomplished by flow cytometry (FCM) analysis, chromosome observation and rDNA analysis.     

CIMMYT's approach to breed for drought tolerance

Summary About 32% of the 99 million ha wheat grown in developing countries experiences varying levels of drought stress. Three major drought types have been identified: Late drought (LD) is common in the Mediterranean region, early drought (ED) is found in Latin America and wheat is produced on residual soil moisture (RM) in the Indian subcontinent and part of Australia. Until 1983, CIMMYT selected all germplasm under near optimum conditions for its yield potential and tested only advanced lines under drought. In spite of many critics, this approach proved to be successful, since in the mid 80's CIMMYT germplasm was grown on 45% of the wheat area in LC with annual rainfall from 300–500 mm and on 21% in areas with less than 300 mm. Since 1983, CIMMYT's drought breeding methodology is to alternate segregating populations between drought stressed and fully irrigated conditions (FI) and to test advanced lines under a line source irrigation system. To compare the efficiency of these approach, yield of four, mostly leading varieties, from each of the regions with LD, ED, RM, and FI and twelve recent CIMMYT cultivars selected for high yield under FI and RM conditions (ALT) were compared under four different moisture regimes (FI, LD, ED, and RM) in 89–90 and 90–91 in Yaqui Valley, Mexico. Genotypic correlation between yield and days to flowering, days to maturity, height, grains m^-2, TKW, test weight and grain fill period were calculated.Mean grain yield of the four best lines in the ALT group was highest under all moisture stress regimes, followed by the FI-group. However, the highest yielding cultivar within each moisture regime was from the FI-group under FI, from the LD-group under LD, and from the ALT-group under ED and RM conditions. Estimates for genetic advance suggest that FI is the best environment for increasing grain yield even in all three drought environments. This indicates that yield potential per se is beneficial also in drought environments. The highest yield in drought environments was realized by the CIM cultivars selected under FI and RM. Simultaneous evaluation of the germplasm under near optimum conditions, to utilize high heritabilities and identify lines with high yield potential, and under stress conditions to preserve alleles for drought tolerance seem at present the best strategy.     

Identification of bread wheat,durum wheat and barley cultivars adapted to dry areas of Southern Italy

Summary Among the cultivars of bread wheat, durum wheat and barley grown in the South of Italy, genetic variation for adaptation to the high temperature and drought stress conditions typical of the Mediterranean environment has been found.The basic data have been extrapolated from 5 years of Italian national network cultivar trials, where 20–30 cultivars were grown in replicated plot trials in 30–50 locations per year, including some where stress strongly affected grain yield.After careful identification of the most representative years and testing sites it was possible to characterise the cultivars on the basis of the grain yield in stress conditions and the Fischer & Maurer (1978) susceptibility index and to find genotypic differences sufficiently repeatable in years.The cultivars giving the best yield under stress associated with low susceptibility indices were in bread wheat: Etruria, Spada, Pandas, Centauro, Oderzo, Costantino and Gladio, in durum wheat: Aldura, Arcangelo, Adamello, Vespro and Capeiti, in barley: Fleuret, Barberousse, Jaidor, Express, Trebbia, Georgie, Dahlia, Criter and Magie.     

An assessment of morphogenic and transformation efficiency in a range of varieties of potato (Solanum tuberosum L.)

Summary To provide a truly genotype-independent transformation system, it is necessary to be able to transform a wide range of potato genotypes. The ability to regenerate shoots in vitro was determined for 34 potato varieties using tuber disc explants. Following a culture regime used extensively in previous studies with the variety Desiree, half of the varieties could be regenerated from tuber discs and half could not. From a sample of varieties that could be regenerated from tuber discs, all but one variety gave transgenic plants. Twelve varieties were evaluated for the capacity to regenerate shoots from leaf and internode explants excised from in vitro grown plants. All of the varieties tested regenerated adventitious shoots. Leaf and internode explants from 5 varieties were subsequently used for transformation, and transgenic plants were produced from two potato varieties that did not give transgenic plants from tuber disc explants. Some varieties could not be transformed by either method, and will require modification of the in vitro regeneration and transformation system to be successful.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid     

Application of image analysis for variety testing of mushroom

Summary A restriction enzyme map of the ribosomal RNA genes (rDNA) in Solanum tuberosum cultivars Golden Wonder and Desiree has been constructed. An heterologous probe pTa 71 containing the rDNA derived from wheat was used to detect and map the corresponding region in potato genomic DNA fragments. rDNA repeats of cultivars Desiree and Golden Wonder are similar with respect to their target sites for restriction enzymes, however, these cultivars have diverged with respect to the length of their intergenic spacer (IGS) sequences. IGS length variants may reflect the different breeding systems for Golden Wonder and Desiree. Furthermore, some of the 25S genes of both cultivars appear to have a 100–150bp insertion/deletion near their 3 end. The presence of RFLPs (restriction fragment length polymorphisms) in the IGS region could be used as a DNA fingerprint for potato cultivar identification; the ability to recognise IGS length variants may be of significance to potato phylogenetics and breeding.     

Genetic analysis of,and selection for,factors affecting quantitative resistance to Xanthomonas campestris pv. oryzae in rice

Summary This study assessed 46 potato cultivars, breeding lines and Solanum spp. for heat-tolerance using short-term growth rates and carbon assimilation measurements of young in-vitro-derived plants. Plants of the 46 clones and species were transferred from greenhouse conditions to controlled conditions set at 33/25°C day/night with 12 h photosynthetic photon flux density (PPFD) at 430–450 mol m^-2s^-1 and an 8 h daylength extension (6 mol m^-2s^-1), to inhibit tuberization. Twenty eight accessions were also grown in a 20/10°C controlled environment. Plants were harvested after 10 and 20 days and dry weights of the plant components were measured for plant growth analysis. Gas exchange (leaf net photosynthesis and maintenance dark respiration) and leaf chlorophyll fluorescence parameters (O, P, and T) were measured at 30°C. Amongst the 30 accessions grown at both hot and cool temperatures, only two accessions (Yungay and AVRDC 1287.19) produced more dry weight in the hot chamber than the cool chamber. Hot/cool ratioss for net assimilation rate (NAR) and relative growth rate (RGR) exceoded unity in five and six accessions, respectively. For the 46 accessions grown under hot conditions, none had significantly greater values than those of the control clones for RGR and NAR. Differences between clones in maintenance respiration and net photosynthesis were more closely related to RGR, NAR, and total dry weight (TDWT) in clones which invested more dry weight in leaves and less in stems. Attributes of the chlorophyll fluorescence curve did not explain more of the clonal variation in RGR, NAR, and TDWT than did gas exchange parameters. No single gas exchange or fluorescence character explained more than 50% of the variability among clones for NAR, RGR, or TDWT, but combination of favourable attributes could improve potato heat tolerance in the future.