Methods for Selection of Low-Temperature Tolerant Mutants of Chrysanthemum morifolium Ramat. Using Irradiated Cell Suspension Cultures II. Preselection in vitro under Low-Temperature Stress

The effect of in vitro cold treatment (+6°C, 90 days) on selection of low-temperature tolerant (LTT) mutants was investigated using X-irradiated (15 Gy and 20 Gy) and non-irradiated cell suspensions of Chrysanthemum morifolium cv. ‘Parliament’. Low temperature reduced significantly the number of regenerating calli, especially in the irradiated cultures. After the end of the cold stress 684, 197 and 54 plants were obtained from the 0 Gy, 15 Gy and 20 Gy treated cultures, respectively. Testing these plants in the greenhouse (+12 °C, winter low-light) most of the early flowering variants were selected from the 15 Gy population.     

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.     

A model describing the flowering of single plants, and the heritability of flowering traits of Dimorphotheca pluvialis

In the development of new crops such as Dimorphoteca pluvialis (L.) Moench, improvement of flowering synchronisation is an important breeding objective. The flowering of single plants of Dimorphotheca pluvialis could be described by a logistic curve obtained by the regression of cumulative number of open flowers on time. The curve is characterised by three parameters, corresponding with the total number of flowers produced by the plant, the rate of flowering development and the day at which peak bloom is reached. From these parameters two other characteristics were derived, i.e., onset of flowering and duration of flowering. The use of the flowering model for selection for improved flowering synchronisation is discussed. Heritabilities of flowering traits were estimated using parent-offspring regression and variance components analyses. Onset of flowering and date of peak bloom showed high (>0.69), and total number of flowers moderate to high (0.30–0.90) heritability values, indicating that for these traits considerable progress may be expected from mass selection, particularly in the early selection generations. Duration of flowering showed low to moderate values (0.25–0.45), and methods other than mass selection (e.g. family selection) should be considered. Determination of phenotypic and genetic correlations revealed only an additive genetic correlation between date of peak bloom and duration of flowering (r _A = 0.80 and 0.69 for 1993 and 1994, respectively), suggesting the possibility of indirect selection for curtailed duration of flowering by means of selection against late date of peak bloom. Duration of flowering, total number of flowers and onset of flowering were not correlated. This revised version was published online in August 2006 with corrections to the Cover Date.     

Soyabean-seed Yield Response in Row- and Hill-plots under a Recurrent Selection Programme

Several studies have indicated a possible link between genome size and earliness in maize. In this study, an original maize population, South African photo-period insensitive maize composite II (C₀) and several selected generations were analyzed for days to flowering, plant height, ear height, and yield. The selection criterion was earliness. Over six cycles of selection a 14-day decrease in days to flowering was obtained with no significant reduction in grain yield. In the initial population (C₀) and the most advanced selection (C₆) 101 plants were analyzed for nuclear DNA content. A significant decrease in the mean nuclear DNA content of the C₀ population (102 AU) was observed with respect to the C₀ population (105 AU). This reduction was the result of a decrease in frequency of plants in the C₆ population with large genome sizes. Therefore it was concluded that selection for earliness resulted in selection against plants with large genome sizes. Hence, the nucleotype of a maize plant can be modified by selection such as early flowering time.     

Assessment of growth and seed oil composition of kenaf (Hibiscus cannabinus L.) germplasm

This study was conducted to evaluate the growth characteristics and fatty acid composition among 15 kenaf mutants derived from the kenaf germplasm C14 and 15 kenaf accessions originating from Russia, India, China, Iran, and Italy. The overall growth performance (plant height, stem diameter, flowering date, leaf, and flower size) of the stem color mutant lines derived from C14 are similar to those of the original variety. However, the flower color mutant lines derived from C14 showed flowering to occur 10 days later when compared with the original variety and showed smaller leaf sizes than the original variety. Late-ripened kenaf accessions (Jinju, Auxu, and Jnagdae) can yield more bio-mass compared with early or medium-maturing germplasm. The late maturity kenaf (Auxu, Jinju, and Jangdae) has a higher oil percentage than the early maturity germplasm. Linoleic, oleic, and palmitic acids were the predominant fatty acids in all kenaf seeds. The stem color mutant lines significantly surpassed the parental means of all saturated fatty acids. In addition, the flower color mutant lines showed broad ranges of variation in oleic acid. The 15 accessions showed a wide range of fatty acid compositions, spanning from 29.75 to 38.30% saturated fatty acids and 61.70 to 70.24% total unsaturated fatty acids, and the late maturity kenaf has a higher linoleic acid percentage than the early maturity germplasm. The flowering period was highly positively (P ≤ 0.01) correlated with the plant height, stem diameter, oil percent, and linolenic acid (C_18:3), and it was significantly negatively (P ≤ 0.01) correlated with stearic acid (C_18:0). These results will provide valuable information to assist the parental selection of kenaf breeding.     

Geneticanalysis of drought stress response in rapeseed (Brassica campestris and B. Napus). II. Yield improvement and the application of selection indices

Summary The expected improvement in grain yield if selection was made for plant characters measured between flowering and maturity was evaluated in populations of rapeseed (Brassica campestris and B. napus) grown in a droughted environment. Drought was commenced at flowering in each species and measurements were made on plants at the commencement of the drought stress, during the stress treatment and at crop maturity.Substantial genetic and phenotypic variation was observed in yield as well as the different morphophysiological determinants of yield. In B. campestris no single parameter was found to be a suitable alternative selection criterion to yield since the correlated responses in yield if selection was for another character was lower than if selection was for yield alone. By the use of a selection index however, joint selection for yield, as well as harvest index, 1000 seed weight and seeds per pod, was expected to be 20% more effective than direct selection for yield under drought. In the B. napus population direct selection for flowering time or for harvest index was predicted to result in a genetic advance in yield equal to or greater than that obtained by direct selection for yield, whereas joint selection for flowering time and yield should result in a 16% greater yield increase. Selection for increased yield in these populations grown in droughted conditions is discussed.     

Genetic analysis in Chrysanthemum morifolium. I. Flowering time and flower number at low and optimum temperature

Summary The number of days to flowering and the number of flowers per plant were studied at 12, 13, 15 and 17 C night temperature in 79 F₁ populations from 15 parents. Only a few F₁ plants flowered earlier than their parents; the majority flowered at the same time, later or not at all. The number of flowers of the F₁ plants was higher than that of the parents.Genetic analyses showed a broad sense heritability of 70% for days to flowering and flower number and a highly significant GCA effect for days to flowering.The progenies and parents that flowered early at one temperature were generally early at all temperatures.     

Over-expression of <Emphasis Type="Italic">Arabidopsis</Emphasis><Emphasis Type="Italic">FT</Emphasis> gene reduces juvenile phase and induces early flowering in ornamental gentian plants

Ornamental gentian plants are perennial and have a juvenile period of over 1 year before flowering. We transformed gentian plants with a construct comprising the Arabidopsis FLOWERING LOCUS T (FT) gene (encoding a major component protein of the flowering hormone ‘florigen’) under the control of the rolC promoter from Agrobacterium rhizogenes, which is known to induce vascular-specific expression. The resultant rolCpro-FT transgenic gentian plants showed early flowering in vitro and the earliest line formed floral buds within 4 months after transformation. Regeneration experiments from leaf explants of these rolCpro-FT transgenic plants also confirmed the early flowering phenotype. After acclimatization, these transgenic plants showed normal floral development in a closed greenhouse. There is no effective method to induce early flowering by cultivation management in gentian, therefore these lines might be very useful as annual early season cultivars.     

Methods for Selection of Low-Temperature Tolerant Mutants of Chrysanthemum morifolium Ramat. Using Irradiated Cell Suspension Cultures III. Comparison of Mutants Selected with or without Preselection in vitro at Low-Temperature

One hundred variants of Chrysanthemum morifolium cv. ‘Parliament’, selected on a single plant basis for early flowering at low temperature, were tested on a clonal basis. These variants originated from irradiated or non-irradiated microcalli subjected to a one-step greenhouse selection procedure, or, a two-step selection procedure (preselection in vitro at 6°C followed by selection in the greenhouse). Clones originating from irradiated calli flowered significantly earlier than the controls. Most of the low-temperature tolerant (LTT) mutants were found among plants obtained by the one-step selection procedure. Variants originating from slowly regenerating calli yielded more LTT mutants as compared to those of fast(er) regenerating calli. We therefore conclude that, irradiation and subsequent selection of plants from slowly regenerating calli considerably increases the efficiency of selecting LTT mutants. The expected advantage of a preselection in vitro at low-temperature could not be confirmed.     

Selection of Semidwarf, Early Maturing and Blast Resistant Mutants after Mutagenic Seed Treatment in Two Locally Adapted Indian Rice Cultivars

The objective of this study was to induce semidwarf, early maturing and blast resistant mutants in two adapted Indian rice cultivars. Seeds of the rice cultivars ‘Madhu Malti’ and ‘Phul Patas 72’ were treated with gamma rays and ethyl methane sulfonate (EMS) as well as a combination of gamma rays and EMS. A total of 96 dwarf (< 75 cm) and semidwarf (< 100 cm) and 104 early flowering/maturing M₂ plants were identified among the 18,060 M₂ plants observed. Sixty-eight blast resistant M₂ plants were also identified. The true mutant status of these plants has yet to be determined in progeny tests. The induction of desired mutant types in locally adapted cultivars could prove to be superior over their introduction from non-adapted foreign materials via backcrossing in that the important agronomic characteristics of adaptation to the local environment are retained in the mutants.     

Flowering time in oil flax can be influenced by microgametophytic selection

The effect of selection for pollen competitive ability in F₁hybrids on days to flowering and some other morphological traits was studied in oil flax. Various intensity of selection on the rate of pollen tube growth was ensured by the change of the time intervals from pollination to the deletion of a part of the style. In the experimental variants the upper half of the style of F₁hybrids was cut off at 40, 60, and 120 minutes, in the control styles being native. It was shown that F₂ population structure essentialy changed for some morphological traits independence of competition intensity between the microgametophytes. The percentage of early flowering genotypes increased with shortening the period from pollination till style excision. In the treatment where styles were cut off 40 min after pollination the frequency of early flowering plants was highest while the frequency of those plants in 120 min treatment approached the control. Moreover, some treatments considerably influenced the phenotypic ratio in the F₂ population for such traits as petal colour and number of side shoots. It is concluded that a positive relationship exists between pollen competitive ability and early flowering in oil flax. The approach based on selection of fast growing pollen tubes that increases the frequency of early flowering plants could be used in oil flax breeding for early ripeness. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of duration from sowing to first flowering in pigeonpea

Duration from sowing to flowering is the most important trait influencing adaptation in pigeon pea (Cajanus cajan), but the inheritance of this trait has not been elucidated clearly. Crosses were made between two early (60 to 70d) and one late (160 to 170d) flowering pigeonpea genotype and F₁, F₂ and BC₁ populations produced. These populations, comprising 60 to 100 parents, 30 F₁, 400F₂ and 40 to 50 BC₁ plants, were grown under natural (mean13.4 hd^-1) and artificially extended (to 15 hd^-1) daylength and duration from sowing to first flowering recorded. Genetic analysis of the segregation ratios, supported by Chi-square tests, indicated that the duration from sowing to flowering in each of the crosses was controlled by two genes assorting independently and with predominantly additive quantitative effects. The segregation patterns were most clearly defined in the 15 hd^-1 daylength. This revised version was published online in July 2006 with corrections to the Cover Date.     

Changes in morphological and physiological traits associated with recurrent selection for grain yield in maize

Summary Experiments were conducted during the growing seasons of 1975 and 1976 to determine changes in morphological and physiological traits associated with recurrent selection for grain yield in maize (Zea mays L.). Four variety hybrids, BSSS(R)CO x BSCB1(R)CO, BSSS(R)C7 x BSCB1(R)C7 [from a reciprocal recurrent selection program involving Iowa Stiff Stalk Synthetic (BSSS) and Iowa Corn Borer Synthetic #1 (BSCB1)], BS12CO x B14A, and BS12C6 x B14A [from a half-sib selection program involving the open-pollinated variety Alph (BS12) and the inbred tester B14A] were grown at 59 300 and 98 800 plants/ha near Ames, Iowa We obtained data on CO₂-exchange rate (an estimate of photosynthetic rate), grain yield, grain-yield components, flowering dates, maturity traits, light interception and use, shelling percentage, harvest index, and various other plant traits.CO₂-exchange rate did not change appreciably with recurrent selection for yield. Grain yield per hectare and per plant were larger for the improved than unimproved hybrids. Grain-yield components did not change significantly with recurrent selection. Kernel weight of BSSS(R)C7 x BSCB1 (R)C7, however, was larger than that of BSSS(R)C0 x BSCB1(R)C0. Pollen-shed-to-silking interval was shorter for the improved than the unimproved hybrids, and grain-filling duration was longer in C7 x C7 than in C₀ x C₀ of the reciprocal recurrent selection program. Furthermore, improved hybrids were characterized by smaller tassels and more upright canopies. Usually, plant traits and leaf-area-related traits were similar for all hybiids.Although dry-matter productivity was similar for all hybrids, those that were improved by recurrent selection produced more grain per unit leaf area and per unit light interception. Also, BS12C6 x B14A was characterized by a higher harvest index than BS12CO x B14A.We concluded that the source (i.e., photosynthetic capacity) was not limiting grain yield in BSSS(R) x BSCB1(R) and BS12. Increased grain yields that resulted from recurrent selection were consequences of longer grain-filling duration for BSSS(R) x BSCB1(R) and increased translocation of photosynthate from source to sink for both BSSS(R) x BSCB1(R) and BS12.Journal Paper J-8953 of the Iowa Agriculture and Home Economics Exp. Stn., Ames, Iowa. Project No. 2152.     

Orychophragmus violaceus, a Potential Edible-oil Crop

Orychophragmus violaceus, a member of the Cruciferae family, has been found to have a high oil quality with high contents of palmitic (14.3 %) and oleic (20.3 %) acids, and lower contents of linolenic (4.8 %) and erucic (0.9 %) acids. Plants of O. violaceus exhibit a high number of branches, pods per plant, and seeds per pod, which contributes to the high yield potential of this plant. Individual selection was made in the original population of O. violaceus, and a few early, disease-tolerant and high-yielding lines were obtained. Intergeneric hybridization was performed between B. napus and O. violaceus and several hybrid plants (F₁) were obtained. After treatment with colchicine, amphidiploid plants developed. O. violaceus shows great potential for becoming an edible oil crop or being used as genetic material in a rapeseed breeding programme.     

Characterization,inheritance, and molecular study of opaque leaf mutant in mungbean (<Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilczek)

Chlorophyll-deficiency mutants are useful as genetic markers and as materials for studying the photosynthesis process. We characterized the inheritance of the gene controlling an opaque leaf (OL) trait in mungbean (Vigna radiata (L.) Wilczek). An F₂ population was developed from the cross between the OL mutant and ‘Berken’, an Australian mungbean cultivar, to study inheritance and molecular tagging. The population was advanced by selfing to produce F₅ lines from which two normal lines and two OL lines were randomly chosen to study chlorophyll content, seed growth and development, and seed cell morphology. The chlorophyll content in opaque leaf was lower than normal and thus was expected to have lower photosynthetic activity. This resulted in yellowish and shrinking pods and seeds within 15 days after flowering, while those from normal plants extended their growth up to 18 days. The cotyledon transfer cells of the OL plants deformed at 12 days and deteriorated at 14 days after flowering. The OL trait was controlled by a single recessive ol gene which was independent from the genes controlling petiole color and growth habit. We used 193 AFLP primer combinations to tag this gene and found that the marker AGG/ATA was linked with the ol gene at a distance of 3.4 cM.     

Exploitation of the late flowering species <Emphasis Type="Italic">Brassica oleracea</Emphasis> L. for the improvement of earliness in <Emphasis Type="Italic">B. napus</Emphasis> L.: an untraditional approach

The oilseed Brassica rapa flowers and matures earlier than B. oleracea, as well as their amphidiploid B. napus. Therefore, earliness of B. rapa has been investigated as a source of variation for earliness in B. napus breeding programs. Variation for days to flower exists in B. oleracea; however, its earliest flowering variant B. alboglabra flowers 2–3 weeks later than B. napus. We hypothesized that the C genome of B. alboglabra carries alleles for early flowering which are different from the C-genome alleles of B. napus; and these alleles can be used for the improvement of B. napus. To test this, we examined flowering time in pedigree and DH populations from two B. napus × B. alboglabra crosses. A B. napus line with about a week earlier flowering than the B. napus parent was achieved through reconstitution of its C genome following pedigree selection. Introgression of the B. alboglabra allele in the early flowering pedigree lines is also evident from the presence of B. alboglabra-specific SSR alleles in this line. However, application of doubled haploidy failed to generate any line that flowered earlier than the B. napus parent, which is probably due to the difficulty of obtaining large numbers of euploid B. napus DH lines from this interspecific cross. Thus, we demonstrate that a trait of the diploid species, which apparently looks undesirable, might in fact be highly valuable for the improvement of amphidiploids; and knowledge from this research can also be applied for other traits.     

Quantitative trait loci for early maturity and their potential in breeding for earliness in Brassica juncea

A doubled haploid population of Brassica juncea, developed from a cross between two parental lines differing for days to maturity, was used to study the efficiency of indirect selection for a primary trait through selection of secondary trait(s) over direct selection for the primary trait when quantitative trait loci information is available for both primary and secondary traits, and applied. Days to maturity was considered as primary trait, while days to first flowering, days to end of flowering, flowering period and plant height were considered as secondary traits. An RFLP linkage map was employed for QTL analysis of maturity and maturity-determinant traits, and a stable QTL B6 simultaneously affecting these two types of traits was identified. This linked QTL explained 11.7% phenotypic variation for days to maturity, 20.7% variation for days to first flowering, 24.3% variation for days to end of flowering and 14.4% variation for plant height. Phenotypic evaluation of maturity and/or maturity-determinant traits, viz. days to first flowering, days to end of flowering and plant height revealed that limited genetic advance for early maturity can be achieved through phenotypic selection of the primary and/or the secondary trait(s). However, the estimates of genetic advance for early maturity based on combined phenotypic evaluation and linked QTL data was found to be, at least, three times higher compared to genetic advance based on phenotypic evaluation only, demonstrating the potential of marker-assisted selection in breeding for early maturity in B. juncea.     

Intergeneric crosses for the transfer of resistance to the beet cyst nematode from Raphanus sativus to Brassica napus

Summary The possibilities to transfer important traits and in particular resistance to the beet cyst nematode (Heterodera schachtii, abbrev. BCN) from Raphanus sativus to Brassica napus were investigated. For these studies B. napus, R. sativus, the bridging hybrid ×Brassicoraphanus (Raparadish) as well as offspring of the cross ×Brassicoraphanus (Raparadish) ×B. napus were used. Reciprocal crosses between B. napus and R. sativus were unsuccessful, also with the use of embryo rescue. Crosses between ×Brassicoraphanus as female parent and B. napus resulted in a large number of F₁ hybrids, whereas the reciprocal cross yielded mainly matromorphic plants. BC₁, BC₂ and BC₃ plants were obtained from backcrosses with B. napus, which was used as the male parent. F₁ hybrids and BC plants showed a large variation for morphology and male and female fertility. Cuttings of some F₁ and BC₁ plants, obtained from crosses involving resistant plants of ×Brassicoraphanus, were found to possess a level of resistance similar to that of the resistant parent. These results and indications for meiotic pairing between chromosomes of genome R with those of the genomes A and/or C suggest that introgression of the BCN-resistance of Raphanus into B. napus may be achieved.     

Cytogenetics of Brassica juncea×Brassica rapa hybrids and patterns of variation in the hybrid derivatives

Interspecific hybridization is an important tool to elucidate intergenomic relationships, transfer characters across species and develop synthetic amphidiploids, and it has been widely applied for improving Brassicas. The objective of the present study was to create genetic variability in Brassica through interspecific hybridization. Crosses between Brassica juncea (AABB, 2n= 36), and Brassica rapa (AA, 2n = 20) vars toria, yellow sarson, and brown sarson were attempted, and the hybrid derivatives were advanced to the F4 generation. Hybrids were obtained from the crosses B. juncea× toria and B. juncea× yellow sarson. The F₁ plants were vigorous and intermediate to the parents in many morphological traits. The meiotic study of AAB hybrids showed 10 II + 8 I in the majority (71.8%) of cells analysed. A maximum of 12 and a minimum of seven bivalents were also observed in a few cells. The occurrence of multivalent associations (trivalents to pentavalents) at diakinesis/metaphase I and a bridge‐fragment configuration at anaphase I were attributed to homoeology between A and B genomes. A high percentage of plants resembling B. juncea was observed in the F₂ generation. Transgressive segregation in both directions was found for plant height, primary branches, main raceme length, siliquae on main raceme, siliqua intensity, seeds per siliqua and seed yield. There were significant differences for the 14 characters in the F₄ derivatives. Moderate to high estimates of phenotypic and genotypic coefficients of variation, broad‐sense heritability, and expected genetic advance were found for seed yield, 1000‐seed weight, siliquae per plant, seeds per siliqua and days to flowering. Intergenomic recombination, reflected as wide variation in the hybrid progenies, permitted the selection of some useful derivatives.     

Cell selection as a possible reason for the specificity of somaclonal variation in pea

A study of regenerants obtained from long-term callus cultures of different pea (Pisum sativum L.) genotypes revealed specificity of in vitro mutagenesis. The specificity was displayed in the preferential generation of the somaclonal variations affecting quantitative and developmental characters. About 60% of regenerated lines obtained from the cultivar Ranny Zeleny carried mutations in the Lf and Sn loci, which control initiation of flowering. An in vitro study of isogenic lines differing at the Lf and Sn loci illustrates the growth advantages of mutant genotypes in tissue culture. These experiments suggest that mutations affecting developmental characters (e.g. mutations in loci which control flowering behaviour) may be expressed in tissue culture and may cause increased adaptation in mutant cells to the in vitro conditions. Rapid propagation of mutant cells during in vitro culture may lead to a higher proportion of quantitative mutations among the regenerated plants.