Productivity of a doubled haploid winter wheat population under heat stress

Breeding of new winter wheat cultivars with good heat tolerance requires better understanding of the genetic background of heat tolerance. In the present work the effect of heat stress on the 6^th day after heading was investigated in a doubled haploid (DH) population arising from a cross between heat-sensitive (Plainsman V) and heat-tolerant (Mv Magma) cultivars. Averaged over the population, heat stress was found to result in a significant reduction in biomass, grain yield and grain number per plant, and in thousand-kernel weight (TKW) and harvest index. High temperature had the greatest effect on the grain yield, with a drop of 36.2% compared with the control. This could be attributed jointly to significant reductions in the TKW of the main ear and in the grain number of the side tillers. The relationship between the yield parameters was confirmed by the positive correlations obtained for the lines in the population. The diverse levels of heat tolerance in the different lines were confirmed by the significant differences in the reduction in the chlorophyll content (SPAD index) of the flag-leaves and in yield parameters. The changes in yield components in stress condition, however, can be still the most effective tools for heat stress evaluation.     

Embryo and endosperm development in wheat (Triticum aestivum L.) kernels subjected to drought stress

The aim of the present work was to reveal the histological alterations triggered in developing wheat kernels by soil drought stress during early seed development resulting in yield losses at harvest. For this purpose, observations were made on the effect of drought stress, applied in a controlled environment from the 5th to the 9th day after pollination, on the kernel morphology, starch content and grain yield of the drought-sensitive Cappelle Desprez and drought-tolerant Plainsman V winter wheat (Triticum aestivum L.) varieties. As a consequence of water withdrawal, there was a decrease in the size of the embryos and the number of A-type starch granules deposited in the endosperm, while the development of aleurone cells and the degradation of the cell layers surrounding the ovule were significantly accelerated in both genotypes. In addition, the number of B-type starch granules per cell was significantly reduced. Drought stress affected the rate of grain filling shortened the grain-filling and ripening period and severely reduced the yield. With respect to the recovery of vegetative tissues, seed set and yield, the drought-tolerant Plainsman V responded significantly better to drought stress than Cappelle Desprez. The reduction in the size of the mature embryos was significantly greater in the sensitive genotype. Compared to Plainsman V, the endosperm cells of Cappelle Desprez accumulated significantly fewer B-type starch granules. In stressed kernels of the tolerant genotype, the accumulation of protein bodies occurred significantly earlier than in the sensitive variety.     

Comparison of the Drought Stress Responses of Tolerant and Sensitive Wheat Cultivars During Grain Filling: Changes in Flag Leaf Photosynthetic Activity,ABA Levels,and Grain Yield

Water status parameters, flag leaf photosynthetic activity, abscisic acid (ABA) levels, grain yield, and storage protein contents were investigated in two drought-tolerant (Triticum aestivum L. cv. MV Emese and cv. Plainsman V) and two drought-sensitive (cvs. GK élet and Cappelle Desprez) wheat genotypes subjected to soil water deficit during grain filling to characterize physiological traits related to yield. The leaf water potential decreased earlier and at a higher rate in the sensitive than in the tolerant cultivars. The net CO₂ assimilation rate (P _N) in flag leaves during water deficit did not display a strict correlation with the drought sensitivity of the genotypes. The photosynthetic activity terminated earliest in the tolerant cv. Emese, and the senescence of flag leaves lasted 7 days longer in the sensitive Cappelle Desprez. Soil drought did not induce characteristic differences between sensitive and tolerant cultivars in chlorophyll a fluorescence parameters of flag leaves during post-anthesis. Changes in the effective quantum yield of PSII (Φ_PSII) and the photochemical quenching (qP) depended on the genotypes and not on the sensitivity of cultivars. In contrast, the levels of ABA in the kernels displayed typical fluctuations in the tolerant and in the sensitive cultivars. Tolerant genotypes exhibited an early maximum in the grain ABA content during drought and the sensitive cultivars maintained high ABA levels in the later stages of grain filling. In contrast with other genotypes, the grain number per ear did not decrease in Plainsman and the gliadin/glutenin ratio was higher than in the control in Emese during drought stress. A possible causal relationship between high ABA levels in the kernels during late stages of grain filling and a decreased grain yield was found in the sensitive cultivars during drought stress.     

Drought,high temperature,and their combination affect ultrastructure of chloroplasts and mitochondria in wheat (Triticum aestivum L.) leaves

Plants experience a number of limiting factors, as drought and heat, which are often coinciding stress factors in natural environment. This study evaluated the changes in mesophyll cell ultrastructure in the leaves of two varieties of winter wheat (Triticum aestivum L.), differing in their drought tolerance, under individual or combined drought and heat treatment. Although the individual stress factors affected leaf ultrastructure, the damaging effect of the combined drought and heat was more pronounced and manifested certain differences between genotypes. Chloroplasts and mitochondria were affected in a variety-specific manner under all adverse treatments. The organelles of the drought-tolerant Katya were better preserved than those in the sensitive variety Sadovo. Leaf ultrastructure can be considered as one of the important characteristics in the evaluation of the drought susceptibility of different wheat varieties.     

The shifting influence of drought and heat stress for crops in northeast Australia

Characterization of drought environment types (ETs) has proven useful for breeding crops for drought‐prone regions. Here, we consider how changes in climate and atmospheric carbon dioxide (CO₂) concentrations will affect drought ET frequencies in sorghum and wheat systems of northeast Australia. We also modify APSIM (the Agricultural Production Systems Simulator) to incorporate extreme heat effects on grain number and weight, and then evaluate changes in the occurrence of heat‐induced yield losses of more than 10%, as well as the co‐occurrence of drought and heat. More than six million simulations spanning representative locations, soil types, management systems, and 33 climate projections led to three key findings. First, the projected frequency of drought decreased slightly for most climate projections for both sorghum and wheat, but for different reasons. In sorghum, warming exacerbated drought stresses by raising the atmospheric vapor pressure deficit and reducing transpiration efficiency (TE), but an increase in TE due to elevated CO₂ more than offset these effects. In wheat, warming reduced drought stress during spring by hastening development through winter and reducing exposure to terminal drought. Elevated CO₂ increased TE but also raised radiation‐use efficiency and overall growth rates and water use, thereby offsetting much of the drought reduction from warming. Second, adding explicit effects of heat on grain number and grain size often switched projected yield impacts from positive to negative. Finally, although average yield losses associated with drought will remain generally higher than that for heat stress for the next half century, the relative importance of heat is steadily growing. This trend, as well as the likely high degree of genetic variability in heat tolerance, suggests that more emphasis on heat tolerance is warranted in breeding programs. At the same time, work on drought tolerance should continue with an emphasis on drought that co‐occurs with extreme heat.     

Heat Stress in Wheat during Reproductive and Grain-Filling Phases

Ambient temperatures have increased since the beginning of the century and are predicted to continue rising under climate change. Such increases in temperature can cause heat stress: a severe threat to wheat production in many countries, particularly when it occurs during reproductive and grain-filling phases. Heat stress reduces plant photosynthetic capacity through metabolic limitations and oxidative damage to chloroplasts, with concomitant reductions in dry matter accumulation and grain yield. Genotypes expressing heat shock proteins are better able to withstand heat stress as they protect proteins from heat-induced damage. Heat tolerance can be improved by selecting and developing wheat genotypes with heat resistance. Wheat pre-breeding and breeding may be based on secondary traits like membrane stability, photosynthetic rate and grain weight under heat stress. Nonetheless, improvement in grain yield under heat stress implies selecting genotypes for grain size and rate of grain filling. Integrating physiological and biotechnological tools with conventional breeding techniques will help to develop wheat varieties with better grain yield under heat stress during reproductive and grain-filling phases. This review discusses the impact of heat stress during reproductive and grain-filling stages of wheat on grain yield and suggests strategies to improve heat stress tolerance in wheat.     

Drought tolerance and proteomics studies of transgenic wheat containing the maize C<Subscript>4</Subscript> phosphoenolpyruvate carboxylase (<Emphasis Type="Italic">PEPC</Emphasis>) gene

Enhancing drought tolerance of crops has been a great challenge in crop improvement. Here, we report the maize phosphoenolpyruvate carboxylase (PEPC) gene was able to confer drought tolerance and increase grain yield in transgenic wheat (Triticum aestivum L.) plants. The improved of drought tolerance was associated with higher levels of proline, soluble sugar, soluble protein, and higher water use efficiency. The transgenic wheat plants had also a more extensive root system as well as increased photosynthetic capacity during stress treatments. The increased grain yield of the transgenic wheat was contributed by improved biomass, larger spike and grain numbers, and heavier 1000-grain weight under drought-stress conditions. Under non-stressed conditions, there were no significant increases in these of the measured traits except for photosynthetic rate when compared with parental wheat. Proteomic research showed that the expression levels of some proteins, including chlorophyll A-B binding protein and pyruvate, phosphate dikinase, which are related to photosynthesis, PAP fibrillin, which is involved in cytoskeleton synthesis, S-adenosylmethionine synthetase, which catalyzes methionine synthesis, were induced in the transgenic wheat under drought stress. Additionally, the expression of glutamine synthetase, which is involved in ammonia assimilation, was induced by drought stress in the wheat. Our study shows that PEPC can improve both stress tolerance and grain yield in wheat, demonstrating the efficacy of PEPC in crop improvement.     

Differential drought-induced modulation of ozone tolerance in winter wheat species

Recent reports challenge the widely accepted idea that drought may offer protection against ozone (O(3)) damage in plants. However, little is known about the impact of drought on the magnitude of O(3) tolerance in winter wheat species. Two winter wheat species with contrasting sensitivity to O(3) (O(3) tolerant, primitive wheat, T. turgidum ssp. durum; O(3) sensitive, modern wheat, T. aestivum L. cv. Xiaoyan 22) were exposed to O(3) (83ppb O(3), 7h d(-1)) and/or drought (42% soil water capacity) from flowering to grain maturity to assess drought-induced modulation of O(3) tolerance. Plant responses to stress treatments were assessed by determining in vivo biochemical parameters, gas exchange, chlorophyll a fluorescence, and grain yield. The primitive wheat demonstrated higher O(3) tolerance than the modern species, with the latter exhibiting higher drought tolerance than the former. This suggested that there was no cross-tolerance of the two stresses when applied separately in these species/cultivars of winter wheat. The primitive wheat lost O(3) tolerance, while the modern species showed improved tolerance to O(3) under combined drought and O(3) exposure. This indicated the existence of differential behaviour of the two wheat species between a single stress and the combination of the two stresses. The observed O(3) tolerance in the two wheat species was related to their magnitude of drought tolerance under a combination of drought and O(3) exposure. The results clearly demonstrate that O(3) tolerance of a drought-sensitive winter wheat species can be completely lost under combined drought and O(3) exposure.     

Effects of homobrassinolide application on plant metabolism and grain yield under irrigated and moisture-stress conditions of two wheat varieties

Homobrassinolide (BR) was applied either as a seed treatment or foliar spray to two contrasting wheat varieties, viz. C306 (drought tolerant) and HD2329 (drought susceptible), to examine its effects on plant metabolism and grain yield under irrigated and moisture-stress/rainfed conditions. BR application resulted in increased relative water content, nitrate reductase activity, chlorophyll content and photosynthesis under both conditions. BR application also improved membrane stability (lower injury). These beneficial effects resulted in higher leaf area, biomass production, grain yield and yield related parameters in the treated plants. All the treatments were significantly better than the untreated control. Generally, 0.05 ppm either as a seed treatment or foliar spray was more effective than the 0.01 ppm treatment. The drought-tolerant genotype C306 showed more response to BR application under moisture-stress/rainfed condition than HD 2329. Increased water uptake, membrane stability and higher carbon dioxide and nitrogen assimilation rates under stress seemed to be related to homobrassinolide-induced drought tolerance.     

Nitrogen Modulates the Effects of Heat,Drought, and Combined Stresses on Photosynthesis,Antioxidant Capacity,Cell Osmoregulation,and Grain Yield in Winter Wheat

Longer and more intense heat and drought stresses will occur in terrestrial ecosystems in the future. Although the effects of individual heat or drought stress on wheat plants have been largely explored, the regulatory effect of nitrogen (N) on winter wheat under heat, drought, and combined stresses and whether N alleviates damage to wheat plants caused by these stresses remain unclear. Therefore, the objective of the present study was to investigate the growth, photosynthesis, antioxidant enzyme and N metabolism-related enzyme activity, cell membrane system, osmoregulatory substance, and yield responses to heat, drought, and combined stresses in wheat plants and to clarify the regulatory effects of N on the growth, physiological and biochemical characteristics, and yield of wheat plants under stress conditions. The results showed that wheat plant exposure to individual heat or drought stress reduced photosynthesis and N metabolism-related enzyme activities and increased antioxidant enzyme activities, electrolyte leakage (EL), and the contents of MDA (malondialdehyde) and O₂^? (superoxide anion). The above parameters showed typical superposition effects under combined stress. Under individual heat or drought stress, wheat plants treated with a medium (N₂) or high (N₃) N supply maintained higher photosynthesis and N metabolism-related enzyme activities than did those treated with a low N supply (N₁). Enhanced heat and drought tolerance in wheat plants under an appropriate N supply may be attributed to improved antioxidant capacity, as exemplified by increased activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), glutathione reductase (GR) and ascorbate peroxidase (APX), and to enhanced osmoregulation capacity, as signified by increased contents of soluble sugar (SS), soluble protein (SP), and proline (Pro). Variable importance in projection (VIP) analysis indicated that efficient SOD, POD, CAT, and GR activities and an increased Pro content had superior potential to alleviate heat, drought, and combined stress stresses in wheat plants, and the improvements in growth and grain yield in wheat plants further confirmed the oxidative stress alleviation and stress tolerance enhancement. However, positive effects of N on wheat growth and grain yield under combined stress were usually observed under a low N supply. These results may facilitate future research on the effects of N fertilizer on the stress resistance of winter wheat.

Graphical Abstract

     

Leaf dehydroascorbate reductase and catalase activity is associated with soil drought tolerance in bread wheat

A number of morphological, physiological and phenological traits have been suggested as significant markers of adaptation to drought in bread wheat (Triticum aestivum L.). This study was aimed at the identification of a relationship between dehydroascorbate reductase (DHAR, EC 1.8.5.1) and catalase (CAT, EC 1.11.1.6) activities in leaves of wheat plants and stability of yield components under water deficit. The single chromosome substitution lines of cv. Chinese Spring carrying separate chromosomes from the donor Synthetic 6x, an artificial hexaploid combining the genomes of the two wild species, Triticum dicoccoides (AABB) and Aegilops tauschii (DD), were the objects of the investigations. The activities of the DHAR and CAT were correlated with flag leaf relative water content and two indexes of stability of grain yield components under drought across the set substitution lines. The lines carrying a synthetic hexaploid homologous pair of chromosomes 1B, 1D, 2D, 3D or 4D all expressed a low constitutive level of DHAR and the lines carrying chromosomes 3B, 1D, 2D and 3D a low constitutive level of CAT. All were able to increase this level (by fourfold for DHAR and by 1.5-fold for CAT) in response to stress caused by water deficit. When challenged by drought stress, these lines tended to be the most effective in retaining the water status of the leaves and preventing the grain yield components from being compromised. The discovered genetic variability for enzymes activity in leaves of wheat might be a useful selection criterion for drought tolerance.     

Genetic dissection of drought and heat‐responsive agronomic traits in wheat

High yield and wide adaptation are principal targets of wheat breeding but are hindered by limited knowledge on genetic basis of agronomic traits and abiotic stress tolerances. In this study, 277 wheat accessions were phenotyped across 30 environments with non‐stress, drought‐stressed, heat‐stressed, and drought‐heat‐stressed treatments and were subjected to genome‐wide association study using 395 681 single nucleotide polymorphisms. We detected 295 associated loci including consistent loci for agronomic traits across different treatments and eurytopic loci for multiple abiotic stress tolerances. A total of 22 loci overlapped with quantitative trait loci identified by biparental quantitative trait loci mapping. Six loci were simultaneously associated with agronomic traits and abiotic stress tolerance, four of which fell within selective sweep regions. Selection in Chinese wheat has increased the frequency of superior marker alleles controlling yield‐related traits in the four loci during past decades, which conversely diminished favourable genetic variation controlling abiotic stress tolerance in the same loci; two promising candidate paralogous genes colocalized with such loci, thereby providing potential targets for studying the molecular mechanism of stress tolerance–productivity trade‐off. These results uncovering promising alleles controlling agronomic traits and/or multiple abiotic stress tolerances, providing insights into heritable covariation between yield and abiotic stress tolerance, will accelerate future efforts for wheat improvement.     

花后干旱或渍水逆境下氮素对小麦籽粒产量和品质的影响

 池栽试验条件下,设置渍水、干旱和对照3个水分处理,每个水分处理下设置两个施氮水平 ,研究了花后渍水或干旱逆境下氮素营养对两个不同类型小麦（Triticum aestivum） 品种籽粒产量和品质性状的影响。结果表明,与对照相比,花后渍水或干旱处理显著降低了小麦的千粒重、穗粒数和籽粒产量。在适宜水分和干旱条件下,增施氮肥增加了小麦籽粒产量,而在渍水条件下,增施氮肥降低了产量。干旱处理提高了蛋白质含量,干、湿面 筋含量,沉降值和降落值;而渍水处理则降低了小麦籽粒蛋白质含量和干、湿面筋含量。同 一水分处理下,增施氮肥提高了蛋白质含量,谷蛋白/醇溶蛋白比,支链淀粉含量和支/直链淀粉比。在小麦籽粒主要品质性状上存在显著的水氮互作效应,且水分、氮肥及水氮互作效 应对小麦籽粒产量和品质的影响因品种的不同而异。     

冀中北小麦品种抗旱性筛选研究

干旱是华北地区小麦生产的重要限制因素,从推广小麦品种中鉴定和筛选抗旱品种,对稳定小麦产量、降低区域水分消耗具有重要意义.本研究以近年来在河北省中北部审定、推广的13个小麦品种为材料,设置冬后浇2水(D2处理)、浇1水(D1处理)和不浇水(D0处理)3种处理方式,进行了大田条件下的抗旱品种筛选试验.结果表明,在D0处理时...     

Variation among wheat (Triticum easativum L.) genotypes in response to the drought stress: I – selection approaches

The agronomic and physiological traits, drought tolerance indexes, principal component analysis and Ward`s method were applied to assess the differences among 20 wheat genotypes in response to drought. Statistically significant correlation was observed for measured traits. Drought susceptibility index (DSI), stress tolerance index (STI) and stress index (SI) were most useful to identify genotypes differing in their response to drought. Utility of the indexes was confirmed by physiological markers of drought tolerance i.e. membrane injury and leaf water status. Variation of the genotypes in biomass and grain yield during drought stress was also verified by clustering methods. Finally, integration of physiological and statistical methods presented in this work, allows to both, indicate that tolerance to drought in wheat has a common genetic background, and select the most diverse genotypes. Based on the results, we recommend a tool for breeders, useful to select the genotypes resistant and sensitive to drought.

Abbreviations: DM: dry matter; DSI: drought susceptibility index; FWC: field water capacity; GY: grain yield; GMP: geometric mean productivity index; H: plant height; LI: leakage index related to membrane injury; M_PRO: mean productivity index; M_HAR: harmonic mean index; NoT: number of tillers; NoG, W-1000: number of grains and weight of 1000 grains, respectively; NoL_MT, NoL_AT, NoL_T: number of leaves on main tiller, adventitious tillers and total leaf number, respectively; PCA: principal component analysis; RTC: relative trait change; RWC, RT, WD: relative water content, relative turgidity and water deficit, respectively SI: stress index; SPAD: leaf greening; STI: stress tolerance index; TI: tolerance index; WCA: Ward`s cluster analysis.     

Root Respiration,Photosynthesis and Grain Yield of Two Spring Wheat in Response to Soil Drying

The effects of soil water regime and wheat cultivar, differing in drought tolerance with respect to root respiration and grain yield, were investigated in a greenhouse experiment. Two spring wheat (Triticum aestivum) cultivars, a drought sensitive (Longchun 8139-2) and drought tolerant (Dingxi 24) were grown in PVC tubes (120 cm in length and 10 cm in diameter) under an automatic rain-shelter. Plants were subjected to three soil moisture regimes: (1) well-watered control (85% field water capacity, FWC); (2) moderate drought stress (50% FWC) and (3) severe drought stress (30% FWC). The aim was to study the influence of root respiration on grain yield under soil drying conditions. In the experiment, severe drought stress significantly (p < 0.05) reduced shoot and root biomass, photosynthesis and root respiration rate for both cultivars, but the extent of the decreases was greater for Dingxi 24 compared to that for Longchun 8139-2. Compared with Dingxi 24, 0.04 and 0.07 mg glucose m⁻² s⁻¹ of additional energy, equivalent to 0.78 and 1.43 J m⁻² s⁻¹, was used for water absorption by Longchun 8139-2 under moderate and severe drought stress, respectively. Although the grain yield of both cultivars decreased with declining soil moisture, loss was greater in Longchun 8139-2 than in Dingxi 24, especially under severe drought stress. The drought tolerance cultivar (Dingxi 24), had a higher biomass and metabolic activity under severe drought stress compared to the sensitive cultivar (Longchun 8139-2), which resulted in further limitation of grain yield. Results show that root respiration, carbohydrates allocation (root:shoot ratio) and grain yield were closely related to soil water status and wheat cultivar. Reductions in root respiration and root biomass under severe soil drying can improve drought tolerant wheat growth and physiological activity during soil drying and improve grain yield, and hence should be advantageous over a drought sensitive cultivar in arid regions.     

Comparative physiological response of wheat genotypes under terminal heat stress

Wheat (Triticum aestivum L.), a staple food crop, is of great commercial importance. Its production is restricted due to multiple environmental stresses. There are indications that the wheat production is consistently limited by terminal heat stress. Previous studies revealed a varied response of different wheat genotypes under heat stress conditions. Here, comparative physiological changes in wheat genotypes viz., DBW-140, Raj-3765, PBW-574, K-0-307 and HS-240 were evaluated under timely and late sown conditions in rabi season. We observed that heat stress dramatically affects chlorophyll content and leaf area index (LAI) in sensitive genotypes whereas proline and malondialdehyde (MDA) content were higher in tolerant genotypes under late sown conditions. Further, the heat susceptibility index (HIS) for 1,000-grain weight, grain weight and grain yield of wheat genotypes viz., HS 240 and K-0-307 was highest as compared with DBW 140, Raj 3765 and PBW 574 genotypes. This finding suggests that wheat genotypes are found to differ in their ability to respond to heat, thereby tolerance, which could be useful as genetic stock to develop wheat tolerant varieties in breeding programs.     

干旱胁迫对大豆生长的影响及抗旱性评价方法与指标筛选

2014-2015年在年降雨量不足40 mm的敦煌市,设干旱和正常灌水两个处理,通过测定12份大豆品种的8个形态指标(株高、主茎节数、有效分枝数、单株荚数、单株粒数、单株粒重、单株生物量和百粒重)及小区产量,采用改进抗旱指数法及权重隶属函数值D对其进行抗旱性评价,并筛选出3份不同抗旱类型的大豆品种进行生理指标测定,验证该方法和指标。结果显示,与灌水处理(CK)相比较,干旱胁迫下的8个形态指标及产量,2014年除有效分枝数及百粒重差异不显著外,其余考察性状均达到了极显著差异,2015年所有考察表型性状均达到极显著差异;两年权重隶属函数法评价结果显著相关,小区产量与两年两种评价方法极显著相关;两种处理下,中度抗旱(中黄24)、弱抗旱品种(WDD00172)的SOD活性、POD活性、CAT活性、丙二醛含量、可溶性糖含量差异极显著,且与权重隶属函数值D显著相关,而强抗旱品种(晋豆21号)中SOD活性、POD活性、脯氨酸含量差异不显著,且6个生理指标与权重隶属函数值D均不相关。因此,权重隶属函数法和小区产量可作为大豆抗旱性评价单一可靠的方法与指标。     

Gene action of canopy temperature in bread wheat under diverse environments

While canopy temperature (CT) shows a strong and reliable association with yield under drought and heat stress and is used in wheat breeding to select for yield, little is known of its genetic control. The objective of this study was to determine the gene action controlling CT in five wheat populations grown in diverse environments (heat, drought, and well-irrigated conditions). CT showed negative phenotypic correlations with grain yield under drought and well-irrigated environments. Additive × additive effects were most prevalent and significant for all crosses and environments. Dominance and dominance × dominance gene actions were also found, though the significance and direction was specific for each environment and genotypic cross. The use of CT as a selection criterion to improve tolerance to drought was supported by its significant association with grain yield and the genotype differences observed between cultivars. Our results indicated that genetic gains for CT in wheat could be achieved through conventional breeding. However, given some dominance and epistatic effects, it would be necessary to delay the selection process until the frequency of heterozygous loci within families is reduced.