Potential of gamma radiation enhancing the biosynthesis of tropane alkaloids in black henbane (Hyoscyamus niger L.)

Summary A productive yellow flowered mutant with high tropane alkaloid content was isolated in the M₂ generation of gamma irradiated (Co⁶⁰ at 40 kr) progenies ofHyoscyamus niger. Mutant progenies, on an average, were capable of synthetising more than twice crude alkaloid (0.147% to 0.221%) in M₃ through M₆ generations compared to the parental control (0.065 to 0.106%). In pilot scale trial (M₇), two elite lines, Y-15 and Y-17 were found to contain 0.390% and 0.318% crude drug in contrast to 0.211% in an improved inbred used as control. However, owing to conducive plant morphology, Y-17 registered high yield of dry biomass, hence gave more than double yield of crude drug than the improved control (inbred) (23 kg/ha against 11 kg/ha). Also being superior in hyoscyamine and hyoscine yields, it was redesignated as var.Aela, and released for commercial cultivation in India, thus signifying the efficiency of radiation breeding for crop improvement.     

Chasmogamous mutant,a novel character enabling commercial hybrid seed production in mungbean

Mungbean (Vigna radiata (L.) Wilczek) is a self-pollinating crop that displays significant hybrid vigor in seed yield of F₁ hybrids. Thus there is the possibility to use hybrid varieties as a breakthrough to raise the yield plateau of mungbean. However, hybrid mungbean seeds can only be accomplished by hand-pollination and thus commercial production is not possible. To encourage hybrid seed set, the plant breeder needs to develop characters that promote higher outcrossing rate such as open flower (chasmogamy). In this experiment, new chasmogamous mutants were induced by gamma irradiation at the rate of 100 and 200 Gy. The mutants were identified at a low rate of 0.4–0.7% in the M₂ generation of accession V1197, and observed for their purity by growing in plant-to-row in the M₃ and M₄ generations. A uniform chasmogamous line was hybridized to normal flower lines to study the inheritance of this character. All F₁ plants had normal flowers, while the F₂ plants segregated well with 3 normal : 1 chasmogamous ratio. When the F₁ was backcrossed to the chasmogamous parent, the progeny gave a ratio of 1 normal to 1 chasmogamous. Thus, chasmogamy was controlled by a single recessive gene, cha.     

Potential of wild germplasm for increasing yield of grain sorghum

Summary Sorghum [Sorghum bicolor (L.) Moench] backcross populations containing 3 to 50% wild germplasm were evaluated in south central India for grain yield and nine related traits. No individual BC₀F₂- to BC₂F₂-derived lines were high transgressive segregates for grain yield. Only 1.5% of all BC₃F₂- or BC₄F₂-derived lines were transgressive segragates, with 26% higher mean grain yield than their respective recurrent parents. The ten highest-yielding BC₂F₂- to BC₄F₂-derived lines per mating having parent CK60B yielded an average of 14% more than CK60B, which was, at the 5% level, a statistically significant difference. However, the increased yield was associated with increased plant height. The highest-yilding lines from RS/R/A2725 x virgatum and RS/R/A2725 x verticilliflorum were an average of 13.5% higher-yielding than RS/R/A2725 (a significant difference) and were equal in plant height. Selection increased BC₂ mean grain yields by 6 to 27%. Population mean yield, mean yield of selected lines, and frequency of high-yielding lines were highest in the BC₄.Journal paper no. 380, ICRISAT, Patancheru, India; Journal paper no. J-11114, Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa, USA.     

Development of high yielding IR64 × <Emphasis Type="Italic">Oryza rufipogon</Emphasis> (Griff.) introgression lines and identification of introgressed alien chromosome segments using SSR markers

Modern rice varieties that ushered in the green revolution brought about dramatic increase in rice production worldwide but at the cost of genetic diversity at the farmers’ fields. The wild species germplasm can be used for broadening the genetic base and improving productivity. Mining of alleles at productivity QTL from related wild species under simultaneous backcrossing and evaluation, accompanied by molecular marker analysis has emerged as an effective plant breeding strategy for utilization of wild species germplasm. In the present study, a limited backcross strategy was used to introgress QTL associated with yield and yield components from Oryza rufipogon (acc. IRGC 105491) to cultivated rice, O. sativa cv IR64. A set of 12 BC₂F₆ progenies, selected from among more than 100 BC₂F₅ progenies were evaluated for yield and yield components. For plant height, days to 50% flowering and tillers/plant, the introgression lines did not show any significant change compared to the recurrent parent IR64. For yield, 9 of the 12 introgression lines showed significantly higher yield (19–38%) than the recurrent parent IR64. Four of these lines originating from a common lineage showed higher yield due to increase in grain weight and another three also from a common lineage showed yield increase due to increase in grain number per panicle. For analyzing the introgression at molecular level all the 12 lines were analyzed for 259 polymorphic SSR markers. Of the total 259 SSR markers analyzed, only 18 (7.0%) showed introgression from O. rufipogon for chromosomes 1, 2, 3, 5, 6 and 11. Graphical genotypes have been prepared for each line and association between the introgression regions and the traits that increased yield is reported. Based on marker trait association it appears that some of the QTL are stable across the environments and genetic backgrounds and can be exploited universally.     

Inheritance of fiber quality and lint yield in a chemically mutated population of cotton

The narrow germplasm base of the upland cotton (Gossypium hirsutum L.), grown on the Texas high plains historically, has limited improvement of fiber quality. Chemical mutagenesis and subsequent selection have helped the development of lines with improved fiber quality in cultivars adapted to this region. This study was conducted to determine the inheritance of improvements in fiber quality. M₃ lines with divergent fiber properties of micronaire, length, and strength were selected from a population of Paymaster HS 200 treated with 3% v/v ethyl methanesulfonate (EMS) for two hours. The 115 selected lines of M₄ and M₅ generation were evaluated for fiber quality and lint yield. Regression of the M₄ and M₅ on the M₃ generation, as well as the M₅ on the M₄ was used to generate narrow sense heritability coefficients. Significant variations were observed between the mutant lines in all generations except for lint yield in the M₅ (1997). The highest heritability estimates were found in fiber length (h ²= 0.29** to 0.46**). Micronaire and strength showed intermediate heritability estimates of h ²= 0.14 to 0.19, while lint yield had a very low heritability estimate of h ²= 0.03. Fiber length and strength were correlated (r= 0.58** to 0.46**) in all the three generations. The mutants identified in these studies have the potential to improve fiber quality of upland cotton without introducing alien genes that may reduce adaptation to short growing season production regions.     

High frequency regeneration and heritable somaclonal variation in Brassica juncea

Summary Multiple shoot formation in cotyledonary callus of Indian mustard (Brassica juncea cv. Prakash) was induced on modified MS media supplemented with high cytokinin (kinetin or zeatin) and low IAA concentrations. Complete plants were obtained on prolonged incubation of shoots on the same medium. 6-Benzyladenine alone or in combination with IAA or NAA did not support plantlet regeneration. A total of 71 plants were transferred to greenhouse. The seed, however, could be collected from 37 plants only. The seed was sown in the field to evaluate the material for somaclonal variation in R₁ generation. Data were recorded for yield, plant height, number of primary branches, siliqua number, 1,000 seed weight and oil content. Somaclonal lines showed tremendous amount of variation for all the characters studied. A number of plants in this generation showed significantly higher yield and/or other improved agriculturally important characteristics as compared to the control. A line with dwarf plant type was also identified. A number of plants were selected from this generation and carried forward to R₂ generation. Most of these lines bred true in R₂ generation. The material seems to be very promising for future breeding programmes.     

Selection for biomass yield in wheat

Summary Biomass (above ground plant parts) yield may be a useful selection trait for yield improvement in wheat (Triticum aestivum L.). This study was conducted to estimate realized heritability of biomass yield and to determine the response to selection for high and low biomass yield in 8 genetically diverse populations of spring wheat under two production systems. Selections were made among the F₃ lines. Progenies of the selected lines were evaluated in replicated field tests in the F₄ generation under high fertility and low fertility production systems at Rampur, Nepal, in 1991. Fertility level had a significant effect on biomass yield, grain yield, effective tiller number, number of kernels per spike, thousand kernel weight, and harvest index. Selection in the F₃ for high and low biomass yield was effective in identifying F₄ lines with high and low biomass yield, respectively. Biomass yield differences between high and low selection groups in the F₄ generation, expressed as percent of the mean of the low selection group and averaged over the eight populations, were 53.9 and 36.5% higher than the mean of the low selection group under the high and the low fertility production systems, respectively. The corresponding figures for grain yield were 48.8 and 34.9% under the high and the low production systems, respectively. Also, selection for high biomass yield resulted in higher effective tiller number, and number of kernels per spike, but lower harvest index. Realized heritability estimates for biomass yield were greater at high fertility (range 0.49 to 0.85) than at low fertility (range 0.22 to 0.44). Biomass yield showed positive genotypic correlations with grain yield, effective tiller number, and number of kernels per spike but a negative correlation with harvest index. The results indicated that selection for high biomass yield should bring about positive improvements in biomass yield, grain yield, effective tiller number, and number of kernels per spike. The correlation between F₃ and F₄ generations suggested that biomass yield in the F₃ generation was a good predictor of biomass yield and grain yield in the F₄ generation. Selection for biomass yield in wheat should be made under the standard production system to obtain a realistic response.     

Honeycomb selection for yield in early generations of spring wheat

Summary The effectiveness of the honeycomb selection method for yield in spring wheat (Triticum aestivum L.) was evaluated using progenies from two wheat crosses, Glenlea x NB131 and Glenlea x Era. Honeycomb selection was carried out in the F₂ and F₃ generations, grown at the University of Manitoba in the summers of 1980 and 1981, respectively. In both generations, divergent selection was made for both high and low yield. Plants selected in the F₃ generation were entered in an F₄ yield test in the summer of 1982. Results of the experiment showed that honeycomb selection for yield in the F₂ and F₃ generations was effective in identifying parents of high- and low-yielding lines. F₃ plants from highyielding F₂ selections gave higher yields than those from low-yielding F₂ selections by 11.5% and 13.0% for Glenlea x NB131 and Glenlea x Era crosses, respectively. The F₄ yield test showed that high yielding selections from both crosses significantly outyielded by 8.9% low yielding selections and by 14.4% the unselected composite lines. It is concluded that the honeycomb selection method can be used for early generation selection in spring wheat.     

Agronomic and morphological characterization of Agrobacterium-transformed Bt rice plants

The agronomic and morphological characteristics of Agrobacterium-transformed rice plants carrying the synthetic cry1Ab or cry1Ac gene were investigated. Tremendous variations in plant height, seed fertility, grain size and other traits were seen in 80 T₁ lines, derived from 80 T₀ plants of 9 rice varieties. On average, about 33% T₁ lines had either morphological or agronomic variant plants. Most of the variations in T₁ plants had no significant correlation with transgene insertion and were proved heritable to their progenies. Genetic analysis in T₃ or T₄ generations showed some simple mutations such as chlorophyll deficiency and stunted plants were independent of transgene insertion and seemed to be controlled by a pair of single genes. However, in two independent transgenic progenies of Xiushui 11, all plants homozygous for transgenes showed dwarfism while all hemizygous and null segregants had normal plant heights. Two advanced homozygous Bt lines, KMD1 and KMD2, were developed from these two progenies. Comparison of the agronomic traits of KMD1 and KMD2 with their parent displayed marked differences among them in terms of seedling growth, tillering ability, yield components and yield potential. The genetic variation observed was generally not linked to the transgene locus and was ascribed to somaclonal variation, but other causes might also exist in particular cases. The results are discussed in the context of choosing appropriate transformation methodology for rice breeding programs. This revised version was published online in August 2006 with corrections to the Cover Date.     

Salt-tolerance in Brassica juncea L. I. In vitro selection,agronomic evaluation and genetic stability

Summary In vitro selection of salt tolerant plants of Brassica juncea L. (Indian mustard) cv. Prakash has been accomplished by screening highly morphogenic cotyledon explant cultures on high NaCl media. Out of a total of 2,620 cotyledons cultured on high salt medium, 3 survived, showed sustained growth and regenerated shoots. They were multiplied by axillary bud culture on NaCl free medium. The salt-selected shoots retained salt tolerance following 3 month of growth and multiplication on control medium. While two of these somaclones flowered and set seeds, third one grew slowly, had abnormal leaf morphology and was sterile. The seed of the two fertile plants were sown in the field to raise R₁ segregating generation. Data were recorded for field, other agronomic components and oil content. The somaclonal lines, both selected salt-tolerant and non-selected, showed tremendous amount of variation for all the characters studied. One of the two tolerant somaclones invariably showed reduced height, longer reproductive phase and higher 1000 seed weight. Based on the agronomic performance of R₁ plants of these somaclones, some plants were selected and their progeny were evaluated for agronomic performance under standard field conditions and salt-tolerance in the greenhouse using sand pot culture method. Most of the lines bred true for their specific characteristics. In the greenhouse, selected salt-tolerant somaclones (SR-2 and SR-3) performed better for plant growth, yield and other agronomic traits at higher salt treatments, indicating thereby that salt-tolerance trait selected in vitro was expressed in the whole plants and is genetically stable and transmitted onto the progeny. The two tolerant lines, however, differed in their salt-tolerance during vegetative and reproductive phases as indicated by their salt-tolerance and stress susceptibility indices. The mechanism of salt-tolerance is not clear and needs to be further investigated.     

Prediction of F6 line performance from F3 families in spring oilseed rape. I. Yield and other agronomic traits

Summary The reliability of a selection among crosses based on a cross prediction in early generations was investigated in spring rapeseed. The performance of the parents, the F₂ generation, and random F₃ lines from four crosses were used to predict the probability of finding superior recombinant lines. These predictions were made for two years and compared with the observed performance of F₆ lines in the second of these two years and in an additional year. Predicted and observed performances coincided reasonably for the characters plant height, standability, maturity and an index calculated from seed yield, oil content and protein content. For seed yield and flowering time, the predictions were very unreliable. In conclusion, prediction methods may be useful in rapeseed breeding, if quality traits are of major commercial interest.     

Fertility improvement in tetraploid pearl millet

Summary Autotetraploid (2n=4x=28) pearl millet inbred lines, Tift 23BE and Tift 239DB, have been developed for use in crosses with other polyploid Pennisetum species. Each line set less than 1% and 3% selfed and open-pollinated seed, respectively. Seed germination was usually less than 17%. First generation (F₁) hybrids between the two inbreds set up to 61% seed while up to 40% of the seed from hybrids germinated.Seed weight per inflorescence for two planting dates averaged 0.32 g and 3.06 g for the two inbreds and the second generation F₂ progeny, respectively. One hundred seed weight was also significantly higher (0.78 g vs 0.48 g) in the F₂ progeny, probably due better endosperm development. Chromosome behavior and pollen stainability were similar in the inbred parents and hybrids. However, the hybrids shed more pollen than the inbred parents.Heterosis was evident in the F₁ hybrids and F₂ progeny which showed significant increases in plant height, leaf length, leaf width, and inflorescence length in addition to fertility improvement.     

Preliminary results of using in vitro axillary and adventitious buds in mutation breeding of Chinese gooseberry

Summary Aseptically cultured shoots of Chinese gooseberry exhibited growth disorder and morphological aberrances, and some died after being exposed to sufficient gamma-ray irradiation. The death rate was dose dependant and the LD₅₀ was 80–90 Gy and 50–60 Gy respectively for cv. Hayward and clone 4. All petiole explants irradiated with gamma-ray could form calli as the control, but the rate of differentiation of adventitious shoots of the petiole explants decreased and was dependant on dose. Sensitivity of the shoot or petiole explants to gamma-ray irradiation varied with species. Gamma-ray irradiation did not deter either the 2-node segments from producing axillary shoots M₁, M₂, and M₃ or the advantitious shoots originating in the petiole explants and the M₃ shoots from forming advantitious roots. Therefore, using aseptically cultured axillary or adventitious buds for mutation breeding of Chinese gooseberry is feasible. A bacterium surviving in the explants lessened the efficiency of these two in vitro techniques in mutation breeding of Chinese gooseberry.Abbreviations IAA 3-indole acetic acid - IBA -indole butyric acid - MS Murashige & Skoog (1962)     

Genotypic variability in vegetable amaranth (Amaranthus tricolor L for foliage yield and its contributing traits over successive cuttings and years

Summary Twenty nine strains of vegetable amaranth (Amaranthus tricolor were grown for two successive seasons to study different selection parameters for foliage yield and its nine contributing morphological and quality traits. The strains AV-38 (5.06 kg/plot) and AV-31 (5.04 kg/plot) recorded highest foliage yield, followed by AV-30 (4.78 kg/plot) and AV-23 (4.70 kg/plot). The protein and carotenoid content averaged 1.24 ± 0.03 mg/100 mg and 0.83 ± 0.02 mg/g respectively. The leaves of A. tricolor also have considerable quantities of ascorbic acid (112.33 ± 5.00 mg/100 g) and fibre (8.39 ± 0.10%). The mean of individual cuttings for plant height, leaf size, stem diameter, foliage yield, protein, ascorbic acid and fibre content increased with successive cuttings till third cutting and thereafter showed a decline. Genotypic coefficient of variation (GCV) values ranged from 6.80 to 28.25%. However, the fibre content, branches/plant, leaves/plant, plant height and stem diameter showed lowest values of GCV. The values of heritability estimates were high for all the traits in all the cuttings as well as on pooled basis and ranged from 0.89 for branches/plant to 0.98 for foliage yield. Highest expected genetic advance was noticed for ascorbic acid (57.48%), followed by foliage yield (48.30%) and leaf size (29.51%).     

Interrelationships among height,lodging and yield in determinate and indeterminate soybeans

Summary Seventy-three determinate and 93 indeterminate random F₉ lines from a cross between determinate and indeterminate soybean [Glycine max (L.) Merr.] parents were evaluated for seed yield, mature plant height, and lodging. Regression analyses indicated that among the determinate lines, seed yield increased by 350 kg/ha and lodging score increased by 0.008 for every 10 cm increase in plant height. Among indeterminate lines seed yield increased by 112 kg/ha and lodging score by 0.3 for every 10 cm increase in plant height. The highest yielding indeterminate lines were tall and very susceptible to lodging. The tall determinate lines had the best combination of high seed yield and excellent lodging resistance.Cooperative Investigations of the Science and Education Administration, U.S. Department of Agriculture and the Purdue University Agricultural Experiment Station.     

Increasing mutagenic efficiency of gamma-radiation in rice by irradiation of seeds at extremely low temperature

Summary Dry seeds of rice, Oryza sativa L., containing about 13 percent moisture, were exposed to ⁶⁰Co gamma raysat extremely low temperature (–196°C). M₁ damage on survival rate, seedling height and seed set were found to become much smaller in the cooling treatment than in the not-cooled control. Chlorophyll and heading date mutation frequencies in the M₂ generation were not affected by the decreased M₁ damage caused by the cooling treatment, and the mutation frequencies could be significantly increased by applying higher doses combined with the cooling treatment. Frequency of sterile or semi-sterile mutants in the M₂ generation was found to be significantly lower in the cooling treatment than in the not-cooled control.It is assumed from the experimental results that extremely low temperatures during irradiation could reduce the amount of physiological damage and gross chromosome aberrations. This results in an increased efficiency of producing minute deletions and/or point mutations.     

Interactions between different genotypic tissues in citrus graft chimeras

The heritability of tolerance of wheat F₃ lines to competition from annual ryegrass (Lolium rigidum), and its relationship to morphological traits, were determined using crosses between wheat cultivars with good and poor competitive ability. Forty F₂-derived F₃ lines from a cross between two late flowering varieties (Machete × Spear) and 40 from a cross between early flowering varieties (Wilgoyne × Gutha) were grown in the field with and without annual ryegrass. There was significant genetic variation between lines within each of the two populations in a number of aspects of plant growth and development, including yield in monoculture. The estimates of heritability for % yield loss due to competition were 0.25 and 0.57 in the two crosses respectively, indicating that selection for high tolerance to competition in the F₃ generation or later should be effective. Fairly strong relationships between height and % yield loss and between leaf length and % yield loss suggest that these may be useful auxiliary traits when selecting for low % yield loss. However, differences between crosses in the magnitude and sign of genetic and phenotypic correlation between traits indicate that competitive ability is a complex character influenced by many factors. This revised version was published online in August 2006 with corrections to the Cover Date.     

Increased grain protein content and its association with agronomic and end-use quality in two hard red spring wheat populations derived from Triticum turgidum L. var. dicoccoides

Suitability of wheat (Triticum aestivum L.) for many food products depends on its unique protein. Elevated grain protein content (GPC) and its quality influences the bread making properties of wheat. The objective of this study was to examine the association of elevated GPC with agronomic and end-use quality in two hard red spring wheat recombinant inbred (RI) populations derived from wild emmer (Triticum turgidum L. var. dicoccoides). The two hard red spring populations (ND683/Bergen and Glupro/Bergen) were developed using a single-seed-descent method. ND683 and ‘Glupro’ are high in GPC (180 g kg^-1), presumably due to the introgression of gene(s) from Triticum turgidum L. var. dicoccoides and ‘Bergen’ is low in GPC (145 g kg^-1). From each of the two populations 12 high- and 12 low-GPC RI lines (F_5:7) were selected for replicated testing at two North Dakota (ND) locations in 1995. In both populations, the high-GPC lines had significantly (p < 0.05) higher values compared to the low-GPC lines for mean GPC and water absorption. Mean grain yield of the high-GPC lines was not significantly different from the low-GPC lines in both populations. In the ND683/Bergen population, the high-GPC lines had significantly (p < 0.05) higher values than the low-GPC lines for mean plant height, days to heading, and flour extraction. GPC was significantly (p < 0.05)and negatively associated with test weight and also significantly (p < 0.01) and positively associated with water absorption in the Glupro/Bergen population. In these populations, results suggested that it may be possible to select lines that combine higher GPC and acceptable yield level, but later in maturity and taller in plant height. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparison of selection procedures in breeding for seed yield in segregating sesame populations

Summary Four hundred sesame (Sesamum indicum L.) lines selected by five methods from five crosses were tested for yield and other agronomic characters in F₆ and F₇. More high yielding lines were isolated by the bulk method and in the cross involving two local cultivars when two seasons means and performance in each individual season were considered. The cross involving two exotic cultivars did not produce a single line superior to local recommended variety. The 80 highest yielding lines recorded a 10.8% increase in yield compared to the mean of 400 lines. Thirty five lines were among the top 20% both seasons with a 13.2% increase in yield. Highly significant seasonal and population effects were recorded. There were no significant differences among the methods of selection although the yield differences among lines were highly significant. Interactions of seasons × populations, seasons × methods, populations × methods and seasons × populations × methods were highly significant. The results indicate that simple and less expensive bulk method may be used with equal success as the pedigree, single seed descent or early generation testing methods when selecting for yield in segregating sesame populations. The importance of inclusion of a high yielding, locally adapted cultivar in the crossing programmes for yield breeding of sesame has also been revealed.Abbreviations BM Bulk method - EGT Early generation testing - G × E Genotype × environment - MSS Modified single seed descent - PM Pedigree method - SSD Single seed descent     

The effectiveness of early generation (F3) yield testing and the single seed descent procedures in two cowpea (Vigna unguiculata (L.) Walp.) crosses

Summary Two breeding procedures were compared in two cowpea crosses. Both procedures were started from the same selected F₂ plants in each cross. For the early generation yield testing, the F₃ lines were yield tested and a pedigree and bulk methods followed in F₄ and F₅. Each F₅ line was bulked to provide seed for a yield test in F₆. In the Single Seed Descent (SSD) procedure, a single seed was taken from each F₂ plant to produce the F₃ generation. The procedure was repeated for the F₄ and F₅. The F₆ SSD lines and the F₆ yield testing lines were compared in yield tests.The results showed that differences in yielding ability of F₃ lines persisted over generations indicating that selection was effective. This was confirmed by the high significant correlations between F₃ yields and those of later generations which ranged from r=0.51^* to 0.85^**.The grain yields of lines derived by the single seed descent procedure were as good as those derived from early generation yield testing.Significant linear correlation between visual rating of F₃ and F₆ yields with actual yields revealed that it is possible to identify promising lines of cowpea visually.