Optimisation of breeding for agronomic traits in fibre hemp (Cannabis sativa L.) by study of parent-offspring relationships

Summary In the framework of a comprehensive Dutch research project the feasibility of hemp cultivation for paper pulp production is being investigated. This project also includes a hemp breeding programme which was initiated at CPRO-DLO in 1990. Hemp breeding is primarily aimed at improving bast fibre production since bast fibre is qualitatively superior to woody core fibre for paper pulp production. The progress of the ongoing breeding programme is hampered by the lack of knowledge on the inheritance of agronomic traits.The following traits were examined on 252 parental plants in 1991 and subsequently on their progenies in 1992: fibre content, plant height, date of flowering, stem diameter and cannabinoid contents. For each trait heritabilities were estimated and direct and indirect effects of artificial selection were studied.The heritability of bast fibre content was high and mass selection proved to be an efficient method causing no undesirable influence on other characters. Characters not directly related to bast fibre yield such as date of flowering, plant height and stem diameter were shown to have disadvantages as selection criteria for the improvement of bast fibre yield.The cannabinoids THC and CBD were studied, as the acceptance of hemp cultivation requires a low level. The content of THC, the psychoactive component can be successfully reduced by mass selection, but it is not certain that mass selection is the most efficient method.     

The inheritance of chemical phenotype in <Emphasis Type="Italic">Cannabis</Emphasis><Emphasis Type="Italic">sativa</Emphasis> L. (IV): cannabinoid-free plants

A genetic factor that blocks the cannabinoid biosynthesis in Cannabis sativa has been investigated. Crosses between cannabinoid-free material and high content, pharmaceutical clones were performed. F₁s were uniform and had cannabinoid contents much lower than the mean parental value. Inbred F₂ progenies segregated into discrete groups: a cannabinoid-free chemotype, a chemotype with relatively low cannabinoid content and one with relatively high content, in a monogenic 1:2:1 ratio. In our model the cannabinoid knockout factor is indicated as a recessive allele o, situated at locus O, which segregates independently from previously presented chemotype loci. The genotype o/o underlies the cannabinoid-free chemotype, O/o is expressed as an intermediate, low content chemotype, and O/O is the genotype of the high content chemotype. The data suggests that locus O governs a reaction in the pathway towards the phenolic cannabinoid precursors. The composition of terpenoids and various other compound classes of cannabinoid-free segregants remains unaffected. Backcrossing produced cannabinoid-free homologues of pharmaceutical production clones with potential applications in pharmacological research. A new variant of the previously presented allele ‘B ₀’, that almost completely obstructs the conversion of CBG into CBD, was also selected from the source population of the cannabinoid knockout factor.     

Cannabinoids for the pharmaceutical industry

Summary Cannabis sativa, is a rich source of a variety of compounds, including cannabinoids, terpenoids and flavonoids. Their content depends upon the plant genetics, growth conditions, time of harvest and drying conditions. To date, more than 60 different cannabinoids have been identified in the plant. Cannabis has been used medicinally for 4000 years and remained in the British pharmacopaeia until 1932, and in the British Pharmaceutical Codex until 1949. Medical use has been prohibited in the UK since 1973. The principal cannabinoid, delta-9-tetrahydrocannabinol (THC) was first isolated in 1964; the first cannabinoid pharmaceutical product Marinol® (a synthetic THC product) was approved in the USA in 1985. The discovery of specific cannabinoid receptors in the early 1990s and subsequent identification of the endocannabinoids anandamide and 2-arachadonoylglycerol, led to a resurgence of interest in the field of cannabinoid medicine, especially within the pharmaceutical industry. Cannabidiol (CBD), as a non-psychoactive, cannabinoid is currently a cannabinoid of significant interest, showing a wide range of pharmacological activity. The other classes of compounds present in cannabis also have their own pharmacology (e.g. terpenoids, flavonoids). The potential for interaction and synergy between compounds within the plant, may play a role in the therapeutic potential of cannabis. This may explain why a cannabis-based medicine using extracts containing multiple cannabinoids, in defined ratios, and other non-cannabinoid fractions, may provide better therapeutic success and be better tolerated than the single synthetic cannabinoid medicines currently available. The development and employment of one of these medicines, Sativex®, is described.     

Correlations within and between characters between and within generations as a measure for the early generation selection in potato breeding

Summary The main objective was to determine the reliability of selection in early generations of a potato breeding programme. To that end data were recorded in the seedling generation, an early and a late harvested experiment of the first clonal generation and the late harvested second clonal generation on up to 30 genotypes of 55 populations from crosses of three Solanum tuberosum ssp. tuberosum cultivars and seven selected S. tuberosum ssp. andigena clones, adapted to the northern temperate zone. Phenotypic correlations were computed per population in almost all possible combinations within and between characters between and within generations. The mean across 55 comparable correlation coefficients was considered to be the best estimate of the relationship concerned.In all four experiments data were collected on number of tubers, tuber yield and mean tuber weight; in the three experiments of the two clonal generations in addition on date of emergence, number of stems, plant height, haulm type, and general impression of the underground plant parts and in the two late harvested clonal generations on date of maturity and under water weight.The results confirmed that general impression which is largely decisive for the selection in early generations, is a little reliable measure. However, from the magnitude of the mean correlations between experiments within characters it must be concluded that selection for most of these separate characters is hardly more reliable. An exception was date of maturity for which a mean correlation was determined of r=0.78 between the two clonal generations. The second strongest mean correlations, perhaps surprisingly, were for tuber yield, viz. r=0.52 between the seedling generation and the late harvested first as well as the second clonal generation and r=0.63 between the late harvested clonal generations. From the correlations between characters it appeared that for a high tuber yield, besides its components, in particular an early emergence, a large plant height, an attractive haulm type and a late maturity are of importance.The main conclusion is that, as long as not enough seed tubers are available for the layout of statistically sound experiments, a negative instead of a positive selection should be applied to avoid the rejection of most of the valuable genotypes in an early stage. On the basis of the results directives for the application of such a negative selection are given.     

A New Approach for Screening Peas (Pisum sativum L.) for Yield Capacity

Efficient nitrogen fixation by legumes has been proved to be important for yield of dry matter. A search was made for easily-measurable characteristics associated with efficient fixation in young pea plants (Pisum sativum L.) which could be used in a selection programme for high yield. Altogether, 17 characteristics were investigated in 120 breeding lines of pea grown in a liquid solution. Of these characteristics, the protein content of the three upper leaves in a 30-day-old plant showed the best correlation with yield capacity. The correlation was also good (r = 0.32—0.94), when samples from the 3 upper leaves were taken in field-grown material. The protein content of the 3 uppermost leaves can be used to reject genotypes that have low initial nitrogen fixation and thereby low yield capacity. This will increase the possibility of testing lines for other agriculturally important characters.     

Breeding transgressive lines of pepper for resistance to Phytophthora capsici in a recurrent selection system

Summary In order to increase the resistance level of pepper (Capsicum annuum) to Phytophthora capsici the main sources of genetic resistance were intercrossed. The parents included 7 varieties from different origins of partial resistance to P. capsici and to other soilborne pathogens. Two cycles of recurrent selection were performed by 7 plant breeders who screened the plants at two developmental stages with various isolates and at two temperatures. Analysis of data revealed a strong influence of the testing conditions on the evaluation of the plants. The level of resistance of the progenies further depended on the pathogen isolate used to screen the plants. Improvement of the whole population was more significant during the first cycle and lines with enhanced level of resistance as compared to the parents i.e. transgressive lines were fixed from the two cycles. These lines were resistant to much higher inoculum concentrations than the original parents and the expression of resistance was stable at high temperature (32°C). These new sources of polygenic resistance were included into a new breeding program to improve the agronomic characters.     

Genetic variation of a safflower germplasm collection grown as a winter crop in southern Spain

Summary A germplasm collection containing 23 accessions of safflower (Carthamus tinctorius L.) was sown in 1993/94 as a winter crop in southern Spain to perform a comparative evaluation. Morphological, physiological and chemical characters were evaluated. Considerable diversity was found in the group. Only one Mexican accession was dwarf (40 cm tall). Most accessions were spiny but there were two spineless Iranian accessions and others with few spines. Variations were also evident for head and seed size. Seed yield per plant varied from 2.5 to 103.5 g and it was correlated with the number of heads, number of branches and plant height. Cultivars with oil content over 35% were identified. Most accessions were of the high linoleic type but there were two with high or intermediate levels of oleic acid, corresponding to the cultivars Ole and Oleic leed. Sowing in November in southern Spain has produced a long growing period (180–240 days) but earliness is common in Indian accessions. A Chinese phenotype with a prolonged rosette stage could be better adapted for cold tolerance. Red flowered spineless cultivars were present in the collection and will be preferred for ornamental or dye purposes.     

Different quantitative trait loci for <Emphasis Type="Italic">Fusarium</Emphasis> resistance in wheat seedlings and adult stage in the Wuhan/Nyubai wheat population

Fusarium head blight (FHB), caused primarily by Fusarium graminearum (Schwabe), is an important wheat disease. In addition to head blight, F. graminearum also causes Fusarium seedling blight (FSB) and produces the mycotoxin deoxynivalenol (DON) in the grain. The objectives of this study were: (1) to compare the relationship between resistance of wheat lines to F. graminearum in the seedlings and spikes and (2) to determine whether the quantitative trait loci (QTL) for FSB were the same as QTLs for FHB resistance and DON level reported for the same population previously (Somers et al. 2003). There was no relationship between FSB infection and FHB index or DON content across the population. A single QTL on chromosome 5B that controlled FSB resistance was identified in the population; the marker WMC75 explained 13.8% of the phenotypic variation for FSB. This value implies that there may be other QTL with minor effects present, but they were not detected in the analysis. Such a QTL on chromosome 5B was not reported previously among the QTLs associated with FHB resistance and DON level in this population. However, because of recombination, some lines in the present study have Fusarium resistance for both seedling and head blight simultaneously. For example, DH line HC 450 had the highest level of resistance to FSB and FHB and was among the ten lines with lowest DON content. This line is a good candidate to be used as a parent for future crosses in breeding for Fusarium seedling resistance, together with breeding for head blight resistance. This approach may be effective in increasing overall plant resistance to Fusarium.     

Honeycomb and simple mass selection for herb yield and inflorescence to leaf + stem ratio in palmarosa grass

Summary Honeycomb and simple mass selection for herb yield and inflorescence to leaf + stem ratio were studied in a population of palmarosa (Cymbopogon martini Stapf. var. motia) grown at a plant spacing of 90 cm. The inflorescence to leaf + stem ratio was studied for indirect selection for oil content. The effects of selection were evaluated at high (30×30 cm) and low (45×45 cm) plant densities. Honeycomb selection was effective in improving herb yield at both plant densities while simple mass selection was effective only at low plant density. Both selection methods were equally effective in improving inflorescence to leaf + stem ratio but only when the populations were grown at low plant density. Increases in oil content were obtained even when selection for an increased inflorescence to leaf + stem ratio was ineffective. Improvements in each of the three studied traits did not affect any other studied trait.     

Variation for resistance to Fusarium head blight in spring barley

Fusarium head blight (FHB) is a fungal disease of barley and other cereals, causing substantial yield and quality losses, mainly due to the contamination of the harvest with mycotoxins. We aimed to evaluate genetic variation for resistance to FHB and its association with other plant characters in diverse barley germplasm in order to identify useful lines for resistance breeding. The 143 barley lines consisted of 88 current European spring barley lines and cultivars, 33 accessions from the genebank at IPK Gatersleben, and 22 lines obtained from North American institutions. We conducted artificially inoculated field experiments with Fusarium graminearum Schwabe during two seasons. FHB severity was evaluated by repeated assessment of visual symptoms. On a set of 49 lines several trichothecene mycotoxins were analyzed. Variation for FHB severity was quantitative. The lines with lowest FHB severity were 'CIho 4196' and 'PI 566203'. Also within the European spring barley collection variation for FHB severity was highly significant. There was a significant negative correlation between plant height and FHB severity (r=– 0.55). FHB severity assessed in the field and the amount of deoxynivalenol in the harvested grains were positively correlated (r= 0.87). Several lines with a useful level of FHB resistance were found or confirmed and are recommended as crossing partners.     

Response of a new IR50/Moroberekan recombinant inbred population of rice (Oryza sativa L.) from an indica × japonica cross for growth and yield traits under aerobic conditions

Genetic variability, correlation, path coefficient analysis and test of normality was conducted in an F₈ recombinant inbred aerobic rice population developed by single seed descent method to evaluate its potential as a mapping population. Estimates of genotypic variance, phenotypic variance, genotypic coefficient of variance (GCV), phenotypic coefficient of variance (PCV), heritability in the broad sense (H) and expected genetic advance at 5% selection index (GA) for grain yield and other attributing characters were computed. In all the cases, PCV was higher than GCV indicating the influence of environment on the characters. High heritability coupled with high GA was observed for several plant traits; number of tillers, plant height, total number of spikelets panicle⁻¹, biomass plant⁻¹, straw weight, harvest index and grain yield plant⁻¹ and hence offered good scope for selection. Grain yield plant⁻¹ was found to be positively correlated with plant height, number of tillers, panicle length, panicle exsertion, number of panicles plant⁻¹, single panicle weight, test weight, number of fertile spikelets panicle⁻¹, straw weight, biomass plant⁻¹, harvest index and grain breadth both at genotypic and phenotypic levels. Harvest index exerted maximum positive direct effect, followed by biomass plant⁻¹ and straw weight on grain yield plant⁻¹ at phenotypic level. Shapiro-Wilks “W test of normality” indicated that the population was skewed towards female parent IR50 for some traits and for some others towards Moroberekan, the male parent. Most of the characters that showed skewness were platykurtic with a kurtosis value of less than 3.     

Genetic combining ability estimates in the F<Subscript>1</Subscript> and F<Subscript>2</Subscript> generations for yield,its component traits and alkaloid content in opium poppy (<Emphasis Type="Italic">Papaver somniferum</Emphasis> L.)

The F ₁ and F ₂ generations of a twenty parent fractional diallel cross of opium poppy (P. somniferum L.) were analyzed for combining ability for ten quantitative and five quality (alkaloids) traits. The results indicated significant differences among the parents for combining ability for all the traits. The GCA and SCA components of variances were significant for all the characters. However, the SCA component of variance (δ ²s) was predominant indicating the preponderance of non-additive gene effect for all the traits except for leaves/plant and papaverine in F₁ hybrids. The average degree of dominance (δ ²s/δ ²g) was more than unity indicating over dominance and also confirming non-additive mode of gene action. Among the parents IS-16, IS-13 and NBRI-1 for early flowering, BR226 and BR241 for branches/plant, capsule weight/plant, seed yield/plant and husk yield/plant, BR227 for leaves/plant, UO1285 for capsule size and opium yield/plant, NBRI-5 for husk yield/plant, morphine, codeine, and thebaine and ‘Papline’ for plant height and papaverine content were found good general combiners. Parent ND1001 was good combiner for codeine and narcotine content. Inclusion of good general combiners in a multiple crossing program or an intermating population involving all the possible crosses among them subjected to bi-parental mating may be expected to offer maximum promise in breeding for higher opium, seed yield and alkaloid contents.     

Recurrent selection for resistance to Verticillium dahliae in pepper

Summary Two cycles of recurrent selection were performed in a population including 7 small fruited varieties of pepper (Capsicum annuum) with low resistance level to Verticillium and resistance to other soilborne pathogens. The collaboration of 7 plant breeders in this work and the use of 2 artificial methods of inoculation allowed screening of numerous plants for resistance to V. dahliae under different conditions. The population was simultaneously improved for resistance to V. dahliae and for resistance to Phytophthora capsici, and at each cycle, intercrosses were performed between all these plants. The general level of resistance to V. dahliae increased continuously during the 2 cycles of seles of selection. Selfed progenies of plants from the 2 cycles displayed a higher resistance than the original parents, whatever the method of inoculation used. This gave evidence of transgressions for resistance. Crosses involving parents that did not seem resistant by themselves also produced transgressive lines, indicating that resistance genes were present but not active in these varieties. The transgressive lines with polygenic resistance to V. dahliae were included into a further breeding program to introduce genes of agronomic value.     

Life history studies of the leafminer Liriomyza trifolii on susceptible and resistant cultivars of Dendranthema grandiflora

Summary The feeding, fecundity, mortality and rate of development of Liriomyza trifolii (Burgess) on 12 cultivars of Dendranthema grandiflora (Tzvel.) (Syn. Chrysanthemum morifolium (Ramat.)) were determined in no choice experiments. Number of feeding punctures, number of eggs laid and total number of larvae varied between cultivars but these characters were not strongly associated with leafminer population increase. Good indicators of host plant resistance were larval mortality, number of pupae and duration of the larval period. Of these three, larval mortality which could be as high as 98% was most suitable.Based on life history data it was predicted that Liriomyza trifolii cannot survive on the resistant cultivar Penny Lane. This prediction was verified experimentally.     

Reciprocal recurrent selection applied to Coffea canephora Pierre

Summary Genetic parameters were estimated in trials planted in connection with a reciprocal recurrent selection programme applied to Coffea canephora in Côte d'Ivoire. Narrow sense heritability values, estimated from a breakdown of the analysis of variance, were high for architectural characters (0.22 to 0.78) and medium for tree vigour (0.13 to 0.40) and for bean weight (0.15 to 0.28). Calculated heritability values for the first two harvests were high (h²>0.7), but much lower for the following harvests (less than 0.2). Heritability estimates by parent-offspring regressions gave variable results, though they were often similar to those obtained by variance analysis, especially for bean weight. Phenotypic and genetic correlations between tree vigour traits and productivity were high. Bean weight was not correlated with vigour or production. Canopy diameter of 4 year-old trees was closely correlated with cumulated productivity from 2 to 5 years in the absence of development competition between trees, but the correlation was low if trees were in competition with each other. The consequences of the results for the choice of characters to be selected among intergroup hybrids or the parents of both populations are discussed.     

Partial resistance in lettuce to downy mildew (Bremia lactucae). 1. Search for partially resistant genotypes and the influence of certain plant characters and environments on the resistance level

Summary In three bicyclic inoculation experiments in the field 756 genotypes from the IVT Lactuca gene bank were screened for differences in partial resistance against Bremia lactucae. Significant differences between genotypes were found. No linkage was found between the presence or absence of R-genes and partial resistance nor was there a clear relationship between plant characters such as size, erectness and lettuce type and resistance level. Comparison of test results for genotypes which were tested twice and in different years and/or environments revealed a rather good relationship. So no clear genotype × environment interactions for resistance level were found. Results imply that the bicyclic inoculation experiments are useful to determine levels of partial resistance.     

Manova and discriminant analyses of phenotypic data as a guide for parent selection in kiwifruit (Actinidia deliciosa) breeding

Evaluation of seedling populations assists in identifying those plants most desirable as parents for a breeding programme. A multivariate analysis of variance (Manova) showed that the seedling populations from 6 kiwifruit (Actinidia deliciosa) crosses differed in their overall merit. Discriminant functions were then used to identify the characters most powerful in distinguishing between populations. The populations were placed in order of their function scores which represent overall merit, and the superior populations were determined. The results showed that `Bruno' was a superior female parent for the production of seedlings with high discriminant function scores for floriferousness (male offspring)and productivity and high vitamin C content (female offspring). One male parent D-1-20 was consistently superior to D-1-6 the other male parent. This revised version was published online in July 2006 with corrections to the Cover Date.     

Analysis of an incomplete diallel cross among three ssp. tuberosum varieties and seven long-day adapted ssp. andigena clones of the potato (Solanum tuberosum L.)

Summary The main aim of the experiment was to study the behaviour of seven long-day adapted clones of Solanum tuberosum ssp. andigena (A) and three varieties of S. tuberosum ssp. tuberosum (T) in an incomplete diallel cross. The four groups of crosses, TxT, TxA, AxT and AxA, held 2, 12, 20 and 21 populations, respectively. Eleven of the intersubspecific crosses were exact reciprocals. Thirty random clones per population were grown as second clonal generation in an incomplete block design with four replications in plots of two plants per clone in 1980, the parents included in many plots. Data were recorded on ten characters.The results showed that group TxA gave the highest tuber yield and AxT the second highest. The F-values indicated highly significant genotypic effects for all characters in all populations. The mean genotypic coefficients of variation (GCV), were the highest for group AxA for all characters. The ANOVA, based on population means showed significant mean squares for general combining abilities (GCA) and for differences of reciprocalls for all characters; the specific combining ability (SCA) mean squares were significant only for date of emergence, plant height and haulm type. Almost all GCA effects of the parents were significant positive or negative. Several populations showed also significant SCA effects. The correlations between the GCA effects and the parental values were of about the same magnitude as the heritabilities; those between the population means and the midparental values were slightly lower.Compared to the midparental values, most populations showed significant positive heterosis for date of emergence, haulm type, number of tubers and under water weight; all populations had significant negative heterosis for mean tuber weight and nearly all for plant height and general impression. All populations from groups TxT and AxA had significant negative heterosis for tuber yield and so had 13 out of the 20 populations from group AxT, while seven of the 12 from group TxA showed significant positive and four significant negative heterosis. The average heterosis for tuber yield of group TxT was –11%, of TxA 3.4%, of AxT–9% and of AxA–18%. The highest mean tuber yields were obtained from crosses with a very high yielding A-parent as female, while the heterosis was negative. Though significant differences between the 11 reciprocals occurred for all characters, consistent differences occurred only for tuber yield. Averaged over the 11 crosses the TxA populations outyielded their exact reciprocals by 10.7%. For the highest tuber yields in A-T crosses, therefore, the tuberosum cytoplasm seems to be preferable. By contrast for a high male fertility the andigena cytoplasm is preferable.     

Effects of recurrent selection for resistance to scab (Gibberella zeae) in wheat

Summary Scab caused by Gibberella zeae Petch., in common wheat, is one of the most severe diseases in China. A source population C0, bred for scab resistance, was developed through three cycles of multiple-parent crossing and intercrossing by means of the dominant male-sterile gene Ta1 (Ms2), according to Wu's scheme. Phenotypic recurrent selection methods for increasing the resistance to scab-infection of spikelets and seeds with the male-sterile plants were carried out simultaneously in Nanjing and Shanghai and at Jianyang, Fujian Province, for three cycles. The generations from C0 to C3 and two check cultivars were evaluated, using a randomized block design, under conditions of an artificially induced epidemic of scab during 1988–1990. The results indicate that there were significant differences in the resistance to scab between these generations. On average, the percentages of diseased spikelets and seeds of the male-fertile plants were reduced by 9% and 10%, respectively. The frequency of resistant plants was distinctly enhanced by recurrent selection. Analysis of variance showed that no significant differences existed between cycles of recurrent selection in agronomic characters such as plant height, spikes per plant, spike length, numbers of spikelets and seeds per spike, weight of seeds per spike and 100-kernel weight, days to heading and to maturity. Except for plant height, most of these traits tended to be slightly improved with improvement of resistance in the gene pool. The variance for resistance in the generations was decreased under selection. Recurrent selection for scab resistance using the dominant male-sterile gene Ta1 (Ms2) was both an effective and feasible breeding method for producing this character in wheat.     

Heterosis breeding in Indian mustard (Brassica juncea L. Czern & Coss): Analysis of component characters contributing to heterosis for yield

Summary Divergence of 25 accessions of Brassica juncea of Indian, CIS (Commonwealth of Independent States, former USSR) and synthetic origin was studied by D² analysis. On the basis of divergence, ten accessions were selected and crossed in a diallel fashion without reciprocals to study the combining ability and heterosis. None of the accessions was found to be a good general combiner for all the nine quantitative characters that were studied. Significant heterosis over better parent for single plant yield was recorded in CIS x Indian and synthetic x CIS crosses (5 each) followed by Indian x synthetic types (3). The analysis of component characters showed that the mean performance of the majority of hybrids was intermediate for five out of six yield attributing traits, thus exhibiting dominance or partial dominance effect. To estimate the contribution of such yield attributing traits towards heterosis for yield, a comparison was made among three parameters viz. heterosis over mid parent (MP), better parent (BP) and better yielding parent (BYP) of the concerned hybrid. It was observed that estimation of heterosis from BYP was a more accurate method to determine the contribution of component characters towards yield heterosis than the analysis based on MP and BP. From the component character analysis, it was concluded that characters like number of primary and secondary branches, number of siliqua per plant and siliqua density contributed significantly towards heterosis in yield. Plot level yield trials of two selected hybrids (Skorospieka II x RH30 and Donskaja IV x Varuna) over two growing seasons revealed 29.4 to 91.8% heterosis over BYP.