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.     

Histochemical and supramolecular studies in determining quality of hemp fibres for textile applications

Summary The composition and supramolecular structure of hemp primary bast fibres have been assessed using microscopy, compositional analysis, wide-angle X-ray diffractometry and CP-MAS ¹³C-NMR, in order to unambiguously define some quality parameters. The main components of the fibre wall were detected by histochemical reactions and modifications occurring during the plant growth have been pointed out. Some differences in fibre lignification degree were recorded among cultivars and confirmed by means of compositional and structural analysis. As for flax and kenaf, X-ray patterns revealed semicrystalline structure of hemp cellulose. NMR spectra and their probabilistic elaboration by MaxEnt method gave further insight on the presence of paracrystalline and amorphous phases and provided an accurate evaluation of polymeric components.     

工业大麻内生真菌菌群结构及其在植株内的空间分布特征

[目的]研究旨在探讨不同样地和不同部位大麻组织中内生真菌的多样性及其分布特征,[方法][结果]从4个不同样地大麻（云麻1号）的植株不同部位组织中共分离得到内生真菌531株,其中茎分离出138株,叶分离出393株。经形态学观察和ITS序列分子鉴定发现其分属于29个属,其中茎22个属,叶24个属。有18个属普遍存在于大麻茎和叶中,其中炭疽菌属是大麻的优势菌属,优势度为40.11%。大麻茎部的内生真菌分离率显著低于叶部,接近地面部分的大麻茎叶组织内生真菌分离率最高。4个样地大麻内生真菌多样性指数在1.84到2.46之间,不同样地中大麻内生真菌组成的相似系数普遍较高,同一样地中大麻茎与叶的相似性系数较小。[结论]结果表明,工业大麻（云麻1号）内生真菌多样性偏低、宿主专一性较小。     

毕节高支苎麻主要农艺性状的遗传分析

对４份毕节高支苎麻的主要农艺性状进行遗传分析的结果表明：鲜皮出麻率和单纤维支数等农艺性状具有较大的遗传力,可在早代单蔸（株）选择或直接选择,产量、鲜皮厚、茎粗等性状的遗传力较低,应根据各性状同相关关系进行间接选择或推迟到晚期世代选择。遗传变幅较大的性状有鲜皮出麻率、株高、单蔸产量等,由于性状的改良效果主要由性状的遗传力和变异幅度及选择强度所决定,因此,在５％的选择强度下,遗传改良效果好的性状为株高     

Histological components of <Emphasis Type="Italic">Trifolium</Emphasis> L. species related to digestive quality of forage

The microscopic analysis of leaflet, petiole, stem and peduncle of 26 populations of 17 Trifolium species was carried out in order to examine their structural characteristics related to digestibility, to compare them between the populations of forage species and their wild relatives and to evaluate, from histological aspect, the potential of wild-growing species to be used as forages. Forage species had low proportion of the tissues composed of thick-walled cells and high proportion of the tissues composed of thin-walled cells in all examined plant parts. The reduction of the proportion of lignified xylem, sclerenchyma and sclerenchymatous interfascicular parenchyma, were suggested as basic selection criteria in breeding Trifolium species for improved digestibility. T. angulatum, T. medium and T. pannonicum proved to be very similar to the forage species in their histological composition, proportion of thick-walled and thin-walled cells and type of variation of the examined parameters. We believe that these species have some potential for use as forages in their own right, but aditional biochemical, physiological and morphological investigations should be conducted.     

QTL mapping of volatile compounds in ripe apples detected by proton transfer reaction-mass spectrometry

The availability of genetic linkage maps enables the detection and analysis of QTLs contributing to quality traits of the genotype. Proton Transfer Reaction Mass Spectrometry (PTR-MS), a relatively novel spectrometric technique, has been applied to measure the headspace composition of the Volatile Organic Compounds (VOCs) emitted by apple fruit genotypes of the progeny ‘Fiesta’ × ‘Discovery’. Fruit samples were characterised by their PTR-MS spectra normalised to total area. QTL analysis for all PTR-MS peaks was carried out and 10 genomic regions associated with the peaks at m/z = 28, 43, 57, 61, 103, 115 and 145 were identified (LOD > 2.5). We show that it is possible to find quantitative trait loci (QTLs) related to PTR-MS characterisation of the headspace composition of single whole apple fruits indicating the presence of a link between molecular characterisation and PTR-MS data. We provide tentative information on the metabolites related to the detected QTLs based on available chemical information. A relation between apple skin colour and peaks related to carbonyl compounds was established. The two authors contributed equally to this work.     

The sexual differentiation of Cannabis sativa L.: A morphological and molecular study

Summary Cannabis sativa L. is a dioecious species with sexual dimorphism occurring in a late stage of plant development. Sex is determined by heteromorphic chromosomes (X and Y): male is the heterogametic sex (XY) and female is the homogametic one (XX). The sexual phenotype of Cannabis often shows some flexibility leading to the differentiation of hermaphrodite flowers or bisexual inflorescences (monoecious phenotype). Sex is considered an important trait for hemp genetic improvement; therefore, the study of the mechanism of sexual differentiation is of paramount interest in hemp research. A morphological and molecular study of Cannabis sativa sexual differentiation has been carried out in the Italian dioecious cultivar Fibranova.Microscopic analysis of male and female apices revealed that their reproductive commitment may occur as soon as the leaves of the fourth node emerge; the genetic expression of male and female apices at this stage has been compared by cDNA-AFLP. A rapid method for the early sex discrimination has been developed, based on the PCR amplification of a male-specific SCAR marker directly from a tissue fragment.Five of the several cDNA-AFLP polymorphic fragments identified have been confirmed to be differentially expressed in male and female apices at the fourth node. Cloning and sequencing revealed that they belong to nine different mRNAs that were all induced in the female apices at this stage. Four out of them showed a high degree of similarity with known sequences: a putative permease, a SMT3-like protein, a putative kinesin and a RAC-GTP binding protein.     

近红外光谱法测定玉米品质指标的研究

应用近红外光谱法(Near infrared spectroscopy,NIRS)和偏最小二乘法(Partial least squares,PLS)建立玉米粗蛋白质、粗脂肪和粗淀粉定量分析的近红外光谱数学模型,并对模型预测结果的准确性进行了评价。结果表明:近红外预测模型的内部交叉验证决定系数(R2cv)分别为:0.9778,0.9666和0.9927;交叉证实标准差(RMSECV)分别为:0.38,0.40和1.51;模型外部验证决定系数(Rv2al)分别为0.9391,0.9651和0.9875;外部验证标准差(RMSEP)为0.41,0.35和1.31。实际样品的常规分析结果得出玉米粗蛋白质、粗脂肪和粗淀粉的NIRS数学模型具有较高的预测准确性,可应用于玉米育种工作中的大批样品的品质分析。     

Quantitative models of hydrolysis conversion efficiency and biomass crystallinity index for plant breeding

In conjunction with improvements to processing technology, selective breeding can be used to increase bioenergy yield potential either through an increase in total harvestable biomass or by modifying biomass conversion efficiency. In this study, the measurement of biomass composition (lignin, cellulose and hemicellulose) and proximate (fixed carbon, volatile matter, ash and moisture contents) components was conducted on leaf and tissue samples from diverse Sorghum bicolor genotypes across two environments. Multiple regression analysis was used to create models that predict two biomass conversion traits: hydrolysis yield potential (HYP) and crystallinity index (CI). CI model variables included tissue type, ash and lignin. HYP model variables included tissue type, ash, cellulose and volatile matter. The presence of ash content in all models was a salient finding of our study. Given that several models were able to predict HYP and CI in alternate grow‐out years, these models may be useful in selective breeding programmes aimed at sorghum bioenergy feedstock improvement through increased conversion efficiency.     

5个工业大麻品种（系）纤维产量及产量构成因素的相关性分析

以自主选育的5个工业大麻品种（系）为研究对象,采用随机区组试验,对纤维产量与产量构成因素进行相关和通径分析,分析参试品种（系）纤维产量、产量构成因素的变化及关系。结果表明,参试工业大麻品种（系）的单株纤维产量及产量构成因素差异显著;单株原茎重与单株纤维产量呈极显著相关,茎粗、单株干茎重、全麻率与单株纤维产量呈显著相关,相关系数的大小依次为单株原茎重（0.962）>全麻率（0.943）>茎粗（0.917）>单株干茎重（0.912）>株高（0.808）>干茎制成率（0.725）;单株纤维产量与单株原茎重、单株干茎重、全麻率、株高、干茎制成率的回归方程达显著水平,通径分析结果表明,对单株纤维产量直接贡献最大的是单株原茎重,其次是全麻率,而株高、单株干茎重、干茎制成率其对单株纤维产量的影响主要是通过影响单株原茎重和全麻率的间接作用而产生的。     

The relationship of stem and seed yields to flowering phenology and sex expression in monoecious hemp (Cannabis sativa L.)

Flowering phenology and sexual dimorphism are two major features that affect stem and seed production in hemp (Cannabis sativa L.), a short-day naturally dioecious plant. The sowing time is of primary importance because it affects flowering time and thereby influences stem yield. In spite of their unstable sexual phenotype, monoecious cultivars facilitate the harvest of both stems and seeds by reducing crop heterogeneity. The main objective of this paper was to determine the stem and seed yields for five monoecious hemp cultivars in relation to their flowering phenology and sex expression. Sowing was carried out on five distinct dates during 2007 and 2008 at two sites in Belgium. The duration from sowing to flowering in days, both stem and seed yields and the seed harvest index decreased when sowing was postponed from mid-April to the end of June. The stem and seed yields from the mid-April sowing (approximately 12.5 and 1.9 t ha⁻¹, respectively) were within the ranges that were reported for fibre and both fibre and seeds production, respectively, in monoecious hemp. No interaction was observed between the sowing date and cultivar for both yields. Sex expression varied among cultivars, indicating that it might be selected, and was partly linked to earliness. Stem yields were lowest in the earliest cultivar (Uso 31) and highest in the latest one (Epsilon 68) while seed yields were lowest in the most masculinized and earliest cultivar (Uso 31) and highest in the most feminized and early (Fedora 17) or mid-early (Felina 32) ones. Both stem and seed yields correlated best with the duration from sowing to full female flowering or from sowing to the end of male flowering.According to our results, harvesting the seeds in addition to the stems in monoecious hemp requires early sowing and the selection of feminized early or mid-early cultivars, earliness depending on the climatic conditions in the cultivation area. Therefore, it might be agriculturally valuable to take sex expression into account in addition to earliness during the selection of cultivars that are adapted to a dual purpose.     

向日葵籽仁脂肪和脂肪酸含量近红外光谱模型的建立

为实现向日葵品质的快速无损检测,选取50份具有代表性的油用向日葵材料,采用偏最小二乘法（PLS）构建籽仁脂肪、亚油酸、油酸、硬脂酸和棕榈酸含量的近红外光谱（NIRS）模型。结果表明,脂肪、亚油酸、油酸含量模型校正和验证相关系数均大于0.96,且预测值与化学值相对误差均在10%以下,能够达到样品成分含量的快速测定。硬脂酸和棕榈酸含量模型校正相关系数分别为0.92和0.82,验证相关系数分别为0.83和0.74,预测值与化学值相对误差在4.66%~17.99%之间,可用于样品成分含量的初步预测。本研究构建的NIRS模型,有助于油用向日葵种质资源品质鉴定和快速筛选。     

Mass selection of wheat for grain filling without transient photosynthesis

Summary Post-anthesis chemical desiccation of wheat (Triticum aestivum L.) plants in the field eliminates transtent photosynthesis by killing all green tissues, thus revealing the plant's capacity for grain filling from stored stem reserves, as the case is for post-anthesis stress such as drought or leaf diseases. This study was conducted to investigate whether mass selection for large kernels under chemical desiccation would lead to the improve ment of grain filling in the absence of transient photosynthesis.Six crosses of common spring wheat were subjected to three cycles of mass selection from F₂ through F₁ when selection was performed for large kernels by sieving grains from plants that were erther chemically desiccated after anthesis, or not (controls). The resulting 36 bulks (six crosses by three selection cycles by two selection environments) were compared with their respective F₂ base populations, when tested with and without chemical desiccation.Selection for large kernels under potential conditions (without chemical desiccation) did not improve kernel weight under potnetial conditions, evidently because these materials were lacking in genetic variation for kernel weight under potential conditions. In four of the crosses, 3rd cycle selection for large kernels under potential conditions decreased kernel weight under chemical desiccation. On the other hand, selection for large kernels under chemical desiccation was effective in improving kernel weight and test weight under chemical desiccation, depending on the cross and the selection cycle, with no genetic shift in mean days to heading or mean plant height. Selection for large kernels under chemical desiccation was also effective in some cases in increasing kernel weight under potential conditions. The results are interpreted to show that selection under potential conditions and under chemical desiccation operate on two different sources for grain filling, namely transient photosynthesis and stem reserve utilization, respectively. In order to expose genetic variability for stem reserve utilization to selection pressure, transient photosynthesis must be eliminated, as done by chemical desiccation in this study.     

Inheritance of early blight resistance in diploid potatoes

Early blight (Alternatia solani) is a fungal disease in hot and humid environments, which causes leaf, stem and tuber lesions. Early blight resistance should be incorporated into potato cultivars because the fungicide spraying is an expensive solution for developing countries. The diploid cultivated species Solanum tuberosum group Phureja and group Stenotomum are sources of resistance alleles. The elucidation of the inheritance for early blight resistance must help to decide what could be the best breeding procedure to improve this diploid germplasm and transfer the resistance to the tetraploid level. Three experiments were carried out under controlled and field conditions to determine the heritability of this trait using nested and diallel mating designs with haploid, species and haploid-species hybrids. The narrow-sense heritability estimates were relatively high (0.64–0.78). This means that additivity was the most important type of gene action for determining resistance to early blight at the diploid level. The results suggested that diploid parents showing highest levels of resistance, throughout the cycle of disease development, can be used in 4x×2x crosses to obtain resistant tetraploid progenies to this fungal disease.     

Isoenzyme and cotyledon protein variation for identification of black beans (Phaseolus vulgaris L.) with similar seed morphology

Summary Methods developed to identify genetically diverse varieties of beans (Phaseolus vulgaris L.) were applied to closely related lines that were difficult to distinguish on the basis of seed morphology. Seedling tissues and seeds of black beans, were examined electrophoretically for isoenzyme and cotyledon proteinn protein patterns. Seven enzymes, extracted from seeds or from seedling stem, root or leaf tissues, were compared for polymorphism. Peptidase, polyphenol oxidase, phosphoglucoisomerase and glutamate oxaloacetate transaminase patterns were the same for all lines. Some differences were observed for acid phosphatase, peroxidase and esterase patterns, but complete discrimination of the six selected lines was not possible on the basis of isoenzyme patterns alone. Using polyacrylamide gel electrophoresis (PAGE) of dissociated 0.1 M acetic acid soluble proteins at pH 3.1 (acid-PAGE), or sodium dodecyl sulphate PAGE (SDS-PAGE) of residual protein extracts, all but one pair of samples in each case could be distinguished from the other samples. Using both techniques all of the lines could be identified unequivocally.     

Exploitation of bacterial pectinolytic strains for improvement of hemp water retting

Summary Retting is the major limitation to an efficient production of textile hemp fibres. Traditional retting has been carried out by autochthonous bacterial community. Aerobic and anaerobic pectinolytic strains were isolated from hemp or flax sources and characterised. Anaerobic pectinolytic strains had a wide range of acid polygalacturonase (PG) activity, whereas aerobic isolates did not produce any acid PG activity, but only an alkalophylic one, suggesting they could play a minor role in the retting process, except in the early stages. Analysis of 16S rDNA sequences assigned anaerobic strains to the Clostridium genus and aerobic isolates to the Bacillus and Paenibacillus genus. C. felsineum and C. acetobutylicum were confirmed as the main anaerobic agents. Nevertheless, a high proportion of anaerobic and aerobic pectinolytic strains was assigned to C. saccharobutylicum and B. pumilus, respectively, both species never being described as involved in water retting. Anaerobic and aerobic strains with high PG activity were selected and characterized. PG activity is well correlated with the strain retting efficiency and improvement of the process was obtained by inoculating the retting water with spores of selected aerobic and anaerobic bacteria. An advisable feature of retting strains is the absence of cellulosolytic activity. An aerobic strain with no cellulosolytic activity was identified. In contrast, all the anaerobic isolates showed cellulosolytic activity. Mutagenesis was ineffective for selection of Cel-Pec⁺ mutants. Localization of the C. felsineum L1/6 PG activity was investigated.     

Effect of Ogura male-sterile cytoplasm on the performance of cabbage hybrid variety. II. Commercial characteristics

Summary The main constraint of using Ogura male-sterile cytoplasm in Brassica is the induction of leaf yellowing at low temperature and the low seed set. But whether or not the cytoplasm can disturb the general commerical performance of the plants is not well-known. Therefore, this work was carried out with the purpose of evaluating Ogura cytoplasmic male-sterile cabbage hybrids and compare them to genomic similar male-fertile ones and to the most popular cabbage hybrid cultivated in Brazil. Harvest data showed an effect of the cytoplasm on reducing plant and head weight, core length and head length and width, although not altering heading index, shape and core indexes, nor head compactness. On the other hand, it was observed also that a careful selection of the parental lineages can produce male-sterile hybrids as good as their male-fertile analogues. All the evaluated hybrids were similar or better than the check for the characteristics under study.Abbreviations dat - days after transplanting - CNPH - National Center for Vegetable Crops Research     

主要麻类作物基因组学与遗传改良:现状与展望

随着测序技术的发展,主要麻类作物(黄麻、红麻、苎麻、亚麻和工业大麻)参考基因组从2011年至2020年陆续完成测序,这标志着麻类作物科学已经进入基因组时代。文章首先详细概述主要麻类作物基因组测序。其次,评述了基于基因组学的麻类作物重要应用价值基因挖掘。基于参考基因组和转录组测序,大量关于纤维发育、响应非生物胁迫的候选基因被挖掘,以促进麻类作物纤维的物种特性和“不与粮食争好地”的逆境农业。同时不同麻类作物特异性状候选基因陆续被报道,如红麻雄性不育、亚麻种子含油量和大麻大麻素相关候选基因等。再次,麻类作物基因组测序完成为基于组学的麻类作物遗传改良提供可能:有助于麻类作物种质资源形成和演化机制研究,系统解析纤维产量、纤维品质、抗病耐逆等农艺性状形成的分子基础;有助于建立高通量基因型-表型数据库,挖掘优异基因资源与创制新种质;有助于创新并集成分子标记辅助选择、基因组选择、转基因等技术,建立高效的快速育种技术体系。宜选育高产高效、抗逆抗病、适宜轻简化机械化、优质专用的多用途麻类作物新品种,以满足麻类作物相关产业的市场需求,适应麻类作物生产方式。尽管已经获得重要基因以及位点的信息,但如何高效率利用已有基因资源对麻类作物进行遗传改良仍需面临一系列挑战,如成熟稳定的遗传转化体系、麻类作物基因编辑体系构建及基因组选择育种等。     

Effect of hardening on frost tolerance and fatty acid composition of leaves and stems of a set of faba bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.) genotypes

Frost tolerance is a main component of winter-hardiness and improving it would promote faba bean (Vicia faba L.) cropping in cool-temperate regions. In many species, leaf fatty acid composition was found to be related to frost tolerance. The objective of this study was to determine, in a representative sample of genotypes, the effect of hardening on leaf and stem (1) frost tolerance and (2) fatty acid composition, and to seek correlations between them. First leaf, second leaf and stem of 31 faba bean genotypes were analyzed after hardening and without hardening. High frost tolerance of known winter genotypes and several experimental lines was shown. Hardening had a significant, positive effect on frost tolerance of all three organs. Stems were on average more frost tolerant than leaves. Hardening induced significant changes in the fatty acid composition: oleic acid decreased significantly in leaves by 3.24% and in stems by 1.77%, whereas linolenic acid increased in leaves by 6.28% and in stems by 9.06%. In stems, correlations between frost tolerance and fatty acid composition were not significant. Correlation coefficients strongly indicated that non-hardened oleic acid content, changes in oleic acid and in linoleic plus linolenic acid content in leaves partly explained their frost tolerance; 0.347 (P < 0.1) < |r| < 0.543 (P < 0.01). The results corroborate the importance of using genetic differences in the fatty acid metabolism in breeding grain legumes for frost tolerance.     

The potential for early generation selection to identify potato clones with resistance to Verticillium wilt

Verticillium wilt (VW) of potato, caused primarily by the fungus Verticillium dahliae, results in yield loss and is therefore an important soil-borne disease. Resistance to VW exists in potato germplasm and is used by breeders during cultivar development. Breeders could make more rapid progress toward the development of VW resistant clones if they had an effective early generation selection strategy. The purpose of this study was to determine whether selection for VW resistance could be carried out in the first tuber generation on single hills. One hundred and fifty-two clones from 19 families were planted as single hills on a V. dahliae-infested field. Each plant was scored for vine maturity, VW symptom expression, yield, stem colonization (colony forming units (cfu), in dried basal stem segments) and incidence (percent infected stems). In the second clonal generation, which consisted of replicated four-hill plots, stem colonization scores and incidence values were used to identify clones which were more resistant than a moderately resistant cultivar and others which were more susceptible than a susceptible cultivar. The efficiency and reliability of the single-hill selection strategy, based on symptoms and yield, was then determined by comparison to the four-hill results. We determined that the best single-hill selection strategy was negative selection (discard clones with the lowest performance) with low stringency, based on yield.