All hermaphrodite progeny are derived by self-pollinating the sunrise papaya mutant

Self‐pollination of a hermaphroditic cultivar normally gives a ratio of 2 : 1 hermaphrodite to female papayas with genotypes M₂m and mm, respectively. Much effort has been dedicated to marking the sexual types of papaya at the seedling stage to distinguish hermaphroditic from female papayas. A hermaphroditic papaya mutant (SR*) has been obtained, derived from the ‘Sunrise’ papaya cultivar mutant. Self‐pollination of the mutant resulted in all progenies being hermaphroditic. The genotype of the female was lethal, as a result of a lethal gene being linked to the mm female gene complex in this case. However, a 3 : 1 segregation ratio was obtained from the progeny of the hermaphroditic cultivar ‘Thailand’ crossed with SR*, indicating that all genotypes survived. Homozygous genotypes (M₂M₂) would be lethal according to Storey's model. Randomly selected F1 plants of the ‘Thailand’ SR* combination were self‐pollinated to obtain an F₂ generation. The F₂ segregation ratio suggested that the SR* mutant had a different form of the M₂ allele, now designated as M_@, which allowed the dominant M_@M₂ to survive in cross combinations. Genetic study has proved that SR* has the M_@ml genotype, a new mutant. It is capable of producing all hermaphroditic papaya progenies.     

The problem of induced sterility: A comparison between EMS and X-rays in Arabidopsis thaliana

Summary The problem of mutagen-induced sterility has been especially studied in barley by Gaul, and in Arabidopsis by Müller and by van der Veen. Their results and conclusions are essentially confirmed and extended by the present Arabidopsis experiment, which compares EMS with X-rays.For a given treatment, M₂-fertility is correlated with M₁-fertility, but mutant frequency in M₂ is equal for the different M₁-fertility classes. It follows that efficiency in mutation breeding can be increased by pre-selection for fertility in M₁, and by using doses which do not make fully fertile M₁-sectors too rare.EMS is more efficient than X-rays by a factor 4 at equal M₁-fertility, and a factor 3 at equal M₂-fertility. As a result of difference in fertility spectrum, fertility improvement from M₁ to M₂ is better for X-rays than for EMS.A considerable load of diplontic sterility and of embryonic lethals (non-germinating seeds) is simply the toll one has to pay for the higher mutant frequency after EMS.Fortunately, part of the diplontic sterility can be pre-selected against in M₁, and maternally conditioned embryonic lethals can be selected against by discarding seedlots from individual M₁-sectors which show reduced germination. (M₂-line selection may also be practised against recessive embryonic lethals and seedling lethals.)     

EMS Induction of Early Flowering Mutants in Spring Rape (Brassica napus)

Studies were undertaken to induce early flowering mutants by ethyl methanesulphonate (EMS) treatments of Brassica napus seeds. EMS treatments for 12 h of a highly inbred B. napus line TBS had adverse effects on M¹ plant development and fertility only when concentrations were greater than 1%. However, an EMS concentration of 1.5% did not reduce M₁ plant fertility to an extent which significantly reduced production of M₂ seeds. Genetic changes induced by EMS treatment and affecting flowering time were of three main types: (1) Changes within a polygenic system reflected by increased variation in flowering time among M₂ families. As the increase in variation was due primarily to a higher frequency of later flowering plants, these polygenic changes would be of little value in developing better-adapted cultivars. (2) Induction of a recessive mutation at a major gene locus which caused M₃ plants homozygous for the mutant gene to flower at least 20 days earlier than the parental line TB8. (3) Induction of a dominant mutation at a major gene locus which affects flowering time by causing a substantial reduction in vernalization requirement. M₂ plants carrying the mutant gene flowered as early as 59 days before the parental line. These major gene mutations could be rapidly exploited in the development of agronomically superior cultivars for short-season, lower rainfall environments in Western Australia.     

Increasing erucic acid content in Ethiopian mustard through mutation breeding

Ethylmethane sulphonate (EMS) was applied to seeds of the Ethiopian mustard (Brassica carinata A. Braun) line C-101. Bulk samples of M₃ seeds from 8331 M₂ plants were evaluated for the fatty acid composition of their oil by near-infrared reflectance spectroscopy (NIRS) and by further gas chromatography on selected samples. A putative mutant, N2-6230, showing very low oleic acid content (4.7% vs. average of 8.6% in C-101) and erucic acid content within the range of variation of the line C-101 (40-49.3%) was identified. The M₃ progeny of this mutant showed a wide segregation for erucic acid content (39.1-57.9% vs. 41.8-50.3% in C-101), and maintained levels of oleic acid lower than in line C-101. Selection for high erucic acid content in the M₃ and M₄ generations led to the fixation of this mutation in the M₅ generation (52.2-59.3% vs. 39.0-47.6% in C-101). This is the first high erucic acid line obtained in Brassica species through mutation breeding. Its utility in future programmes to develop very high erucic acid lines is discussed.     

Radiation and genetical studies to estimate the number of embryonic initials taking part in the formation of the shoot meristem of Vicia ervilia and Lathyrus sativus

Summary Dry seeds of V. ervilia and L. sativus were irradiated with 60Co gamma rays. The M₁ plants were grown to maturity. During harvest, ten randomly selected plants were arbitrarily numbered (symbols P₁ to P₁₀) and the branches and the pod positions along each branch of each plant were numbered (symbols for branches, B₁ to B₄ and pod positions, p₁ to p₁) from the lowest to the topmost one. The seeds of each pod position were collected, threshed, counted and, the next year, sown separately. Mutants were isolated from among the M₂ population and each mutant could be traced back to the particular topography of the M₁ plant. The seeds from the M₂ plants originating from different sectors of the M₁ plants were grown to M₃ generation to study the pattern of inheritance. The segregation data of L. sativus show that in some cases at least one and in other cases at least two embryonic initials were involved in the formation of the shoot meristem whereas in V. ervilia the number of embryonic initials involved was at least two or three.     

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.     

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.     

Dwarf-twisted: An induced mutation in Triticum durum Desf

Summary A mutant phenotype, mainly characterized by undulated leaf surface, dwarfism and sterility, has been repeatedly induced by physical (X-rays; thermal and fast neutrons) and chemical (diethylsulphate and ethylmethanesulphonate) mutagenic agents in different varieties of durum wheat. This phenotype is easily detectable at the seedling stage. Neutron treatments are the most effective method to induce this phenotype and a maximum of 0.6% mutations per M₁ spike-progeny was induced after the highest dose of thermal neutrons. On the average 0.23 mutations per 100 M₁ spike progenies and 0.55 mutant seedlings per 1000 M₂ plants were found after all treatments.Genetic analysis of different dwarf-twisted phenotypes shows that this character is conditioned by one recessive gene with an average segregation ratio of 22.9%. As the dwarf-twisted phenotypes segregate in highly fertile M₁ spikes and no evident chromosome changes have been detected in this caryotype, the hypothesis of a point mutation has been proposed.Contribution no. 171 from the Laboratorio Applicazioni in Agricoltura del C.N.E.N., C.S.N. Casaccia, S. Maria di Galeria, Roma, Italy.     

Heavy-ion-induced mutants in sweet pepper isolated by M1 plant selection

We examined the effects of heavy-ion bombardment on mutagenesis in sweet pepper (Capsicum annuum L.). Dose–response studies indicated that 10 Gy irradiation of ¹²C or ²⁰Ne ions on dry seeds is suitable for inducing mutations in plants. From ²⁰Ne-irradiated M₁ plants, putative mutants included two dwarf plants and one plant whose pericarp was yellow were isolated. Phenotypes of their M₂ progeny were similar to those of the M₁ plants and did not segregate. F₁ plants resulting from reciprocal crosses between the mutants and wild-type plants showed the wild-type phenotype, but phenotypes of F₂ and BC₁F₁ segregated at 1:3 (mutant:wild) and 1:1, respectively. These crossing experiments indicate that the three mutants have monogenic recessive mutations in nuclear genes. In light of these data, we discuss the effectiveness of using heavy-ion bombardment to mutate sweet peppers.     

Somatic Embryogenesis in Maize and Comparison of Genetic Variability Induced by Gamma Radiation and Tissue Culture Techniques

Sixteen inbred lines and one hybrid of manse were tested for their capability of somatic embryogenesis, and fully developed plants could be regenerated, from ten inbred, lines. The highest frequency of plant regeneration was expressed in the inbred line CHI 31, and when this line was crossed with a recalcitrant, non-regenerating line, the F₁ and BC hybrids were regenerable. The results of reciprocal crosses demonstrated that dominant nuclear genes and cytoplasmic factors are primarily responsible for the heritable determination of embryogenic callus proliferation and in vitro regeneration of maize plants. Somaclonal and radiation-induced variability was studied in maize to assess their nature and potential contribution to plant breeding., The inbred line CHI 31 possessing a high in vitro capacity of somatic embryo formation was used as experiments.] material. CHI 31 plants were selfed and twelve-day old zygotic embryos irradiated with ⁶⁰Co gamma radiation in situ. Mature caryopses were harvested and assigned as M₁ material. In another series, immature zygotic embryos (size 1.2—1.5 mm) were cultured in vitro on N-6 medium supplemented with 2,4-D (2.5 μM), and somatic embryos regenerated into plants; these were transplanted into soil and self-pollinated. Regenerants from non-irradiated cultures were grown as R₁ generation, while regenerants from irradiated explants were considered as M₁R₁ generation. The genetic variability was evaluated in the M₂, R₂ and M₂R₂ generations, respectively, and compared with a non-treated seed control. Irradiation induced a variety of chlorophyll and morphological variants segregating in the M; generation; however, the frequency of deviant types obtained in the R: generation (somaclonal variation) was significantly exceeding the one derived from the M₂ populations. The combination of expert irradiation and in vitro regeneration was most effective for the manifestation of chlorophyll and morphological o if types in the M₂R₂ generation, and increased drastically the frequency of early flowering variants. Differences in the segregation patterns of mutant phenotypes amonsister somaclones in the R₃ and M₃R₃ generations indicate a different genetic basis, of plants originating from the same explant. This phenomenon suggests a mutational sectoring of the callus during culture. Radiation induced and somaclonal variation exerted a similar spectrum of chlorophyll and morphological deviants.     

Broad-few-leaflets and outwardly curved wings: two new mutants of chickpea

This study was aimed at the induction of morphological mutations for increasing genetic variability and making available additional genetic markers for linkage studies in chickpea (Cicer arietinum L.). A wilt‐resistant, well‐adapted chickpea cultivar of central India,‘JG 315’(Jawahar gram 315), was used for the induction of mutations. Seeds presoaked in distilled water for 2 h were treated with ethyl methane sulphonate (EMS) using six different concentrations (0.1, 0.2, 0.3, 0.4, 0.5 and 0.6%) and two different durations (6 and 8 h). Several morphological mutants were identified in M₂. One of the mutants, isolated from a treatment of 0.3% EMS for 8 h, had five to nine large leaflets per leaf in comparison with 11‐17 normal‐sized leaflets per leaf observed in the parental cultivar ‘JG 315′. The mutant was designated broad‐few‐leaflets. Many leaves of this mutant showed a cluster of three to five overlapping leaflets at the terminal end. The other mutant, designated outwardly curved wings, was isolated from the 0.5% EMS treatment for 6 h. In this mutant, the wings were curved outwards, exposing the keel petal, while the wings in typical chickpea flowers are incurved and enclose the keel. The lines developed from the broad‐few‐leaflets and outwardly curved wings mutants were named JGM 4 (Jawahar gram mutant 4) and JGM 5, respectively. Inheritance studies indicated that each of these mutant traits is governed by a single recessive gene. The gene for broad‐few‐leaflets was designated bfl and the gene for outwardly curved wings was designated ocw. The locus bfl was found to be linked with the locus lg (light green foliage) with a map distance of 18.7 ± 6.3 cM.     

Effect of gamma radiation on growth and lignin content in Brachypodium distachyon

Brachypodium distachyon has been highlighted as a model monocot plant with small genome and short life cycle. Biofuels are being developed as renewable energy sources to replace fossil fuels. Bioethanol production is negatively correlated with lignin content. Here, Brachypodium was acutely or chronically irradiated at doses of 50, 100, 150, 200, and 250 Gy. The effect of radiation on plant growth and generation of mutant populations was explored. The lethal effect of radiation was higher in acutely irradiated M0 populations. A dose-dependent negative effect in plant height, tiller number, floral spikelet, and total seed number was observed, with a positive effect in days to heading. The phenotype of 1,773 M₁ plants was evaluated, with 417 plants being selected to construct the M₂ population. The 31 M₂ plants that showed the least staining with phloroglucinol were selected. These mutants could be useful materials for studies such as identification of nucleotide substitutions in genes involved in lignin biosynthesis pathway, monitoring of mutant physiological traits, and evaluation of fitness for bioethanol production. As biological resources, the M₂ populations generated in this work will contribute to studies of functional genomics of Brachypodium and to the breeding of grass crops.     

Genetic Analysis of Some Chemotypes in Ocimum basilicum var. glabratum

Screening of a population of Ocimum basilicum var. glabratum Benth., raised from seeds collected from France, West Africa and Kerala (India) was carried out for chemical composition of its essential oils. In all three collections, plants were found to be comprised of 3 chemotypes which were morphologically indistinguishable, but each chemotype was found to be capable of intercrossing freely with the other two. Selfing was carried out during 4 consecutive generations, after which three pure lines were established. Study of the selfed progeny of each collection exhibited an interesting segregation in S₂ and S₃ generations. Along with segregation in F₂ and F₃ generations of various hybrids the results can be explained by assuming a gene ‘M’ existing in 3 (or more) allelic forms: The allele M₀ for biosynthesis of estragole (methyl chavicol) is dominant to allelcs M₁ , for eugenol and M₂ for camphor. Also, allele M₁ is dominant to allele M₂ . The assumption was further confirmed by testcrosses of F₁ hybrids of each cross with their recessive parent.     

Induced mutagenesis for the development of multiline cultivars in bread wheat

Summary Variation for resistance toPuccinia graminis f.sp.tritici, P. recondita f.sp.tritici andP. striiformis was induced in theTriticum aestivum cultivar Lalbahadur using nitrosomethyl urea. Variations were isolated from the M₂ population in the post-seedling stage in the field when infected with a mixture of races of each of the three rusts. Plants exhibiting simultaneous resistance to stem rust, leaf rust and yellow rust were indentified. Repeated screening in the subsequent generations confirmed the resistance of the mutant lines that are morphologically similar to the parental cultivar. The rust resistance of 20 mutant lines was also confirmed at the seedling stage using individual races of stem rust and leaf rust. The different patterns observed in the mutant lines tested against a wide range of races show that these lines can be used as components of a multiline. The patterns of variation compared with those of the known genes for resistance against the Indian races of the pathogens suggest that the mutations for rust resistance are due to factor different from those already known in bread wheat, providing a broadened genetic base for future breeding programmes.     

Breeding for Modified Fatty Acid Composition by Induced Mutations in Linseed (Linum usitatissimum L.)

Seeds of linseed (Linum usitatissimum L.) cultivars ‘Raulinus’ and ‘Bionda’ were treated mutagenically with ethyl methanesulfonate (EMS) in order to broaden the variation of fatty acid composition in the seed oil and to select mutants with reduced linolenic acid (C18: 3) content. The M₂ generation was evaluated for fatty acid composition of the oil. Genetic variation for unsaturated fatty acids was demonstrated to be extended in the M₂ generation as compared to the controls. Finally in the M₅ generation of cv. ‘Raulinus’, lines with reduced C18:3 (< 40% vs. 55%) and correspondingly increased linoleic or oleic acid contents, respectively, could be selected. These mutants proved to be genetically stable as demonstrated by parent-offspring (M₄/M₅) regression.     

The mutagenic effect of ethylene imine (EI) on the tomato

Air-dry tomato seeds were treated with an ethylene imine dilution series. Most sterile M₁-plants can be detected morphologically at an early stage. Few of the morphological deviants proved to be fertile. Both with regard to M₁ ^- and to M₂ ^- growing space the highest concentration leaving fertiles (0.25%) was the most efficient. At this dose the rate of mutation which can be scored on young M₂-plants, reached a value of 1.04 per M₂-family. A small shortage of M₂-families with 1 mutation is suggested by different tests. This may be due to penetrance heterogeneity. The spectrum of chlorophyll mutations changes with the EI concentration, a significantly smaller proportion of chlorodefectios (albinas, etc.) being found at higher concentrations.     

Gamma Ray-Induced Field Tolerance to Phytophthora Blight in Sesame

Due to the non-availability of sources of resistance in Sri Lanka to Phytopbthora blight in sesame (Sesamum indicum) caused by Phytopbthora nicotianae var. parasitica, a mutation breeding programme was initiated. Seeds of three genotypes were subjected to six doses of gamma rays from 100 Gy to 750 Gy from a ⁶⁰Co source. Seeds of M₂ bulks, sampled by variety and treatment from the first five capsules formed on M₁ plants, were grown in a field with a history of repeated incidence of the disease. The best 21 lines, having the highest survival and seed production, identified in the screening of M₃ and M₄ progeny rows of selected M₂ single plants were then tested in a replicated field trial with the recommended cultivar ‘MI 3’ as a control. The plant survival in selected lines averaged 43.3 percent as against 7.2 percent in the recommended variety. Eight selections recorded significantly higher seed yield than ‘MI 3’ at P < 0.01 and another three at P < 0.05. Gamma ray treatments of 450 Gy and 600 Gy produced more lines tolerant to the disease than the other doses used.     

Induction and selection of mutations for resistance against bacterial leaf blight in rice

Summary Dry seed lots of a rice cultivar, Harebare, susceptible for bacterial leaf blight (BLB), were exposed to thermal neutrons with and without pre-treatment of seed for boron-enrichment, and to gamma-rays. M₁ plants of each of the treatments were grown and their seeds were separately harvested. M₂ populations were raised in rice fields of a farmer in a region where BLB is epidemic every year, and M₂ and control plants which expressed resistant reactions to BLB were selected. M₃ and control lines as plant progenies of the selected M₂ plants were raised in the same rice fields as in M₂ generation in order to investigate their BLB resistance and other agronomically important traits. Variances for disease severity of M₂ populations were significantly larger than those of the control population. whereas their means were not singificantly different from that of the control population, suggesting induction of mutations toward both resistance and susceptibility to BLB. Mean disease severity values of the M₃ lines from selected M₂ plants were significantly smaller than those of the control lines, indicating gains of selection in M₂ for quantitative resistance against BLB. Thermal neutrons, especially with boron-enrichment pre-treatment were effective to induce mutations for resistance against BLB. Some M₃ mutant lines with quantitatively enhanced resistance against BLB were not modified in other agronomic traits from those of the original cultivar. Significance of the induced quantitative resistance in breeding programmes for BLB resistance is discussed.     

Selection of 5-methyltryptophan resistant rice mutants from irradiated calli derived from embryos

For increasing the contents of specificfree amino acids in rice (Oryzasativa L.) cultivar, Donganbyeo, mutantcell lines resistant to growth inhibitionby 5-methyltryptophan (5MT) were selectedfrom embryo cultured callus irradiated with50 Gy of gamma rays. Two lines, M₂-1and M₂-2, were obtained from theregenerated plants by selfing. Thesegregation ratios of resistance andsensitivity in these lines fitted 9:7 and3:1 ratios, respectively. Considering theagronomic traits, M₂-1 was about thesame as the original variety in culm lengthand a little longer or higher in paniclelength and number of tillers. However, thefertility of M₂-1 significantlyincreased compared to the originalvariety. M₂-2 had an extremely highculm length and tillering capacity. Four5MT-resistant homozygous M₃lines (MRclasses), M₃-1-40 and M₃-1-116from M₂-1 (MR I),and M₃-2-8 and M₃-2-12from M₂-2 (MR II), were obtained inthe same manner as with the M₂generation. Protein contents of brown ricewere increased about 19% and 32% and the total contents of 9 free essentialamino acids were 71% and 34% greater thanthe original variety in the MR I andMR II groups, respectively. However, forthe free essential amino acids of polishedrice, there was no difference from theoriginal variety.     

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.