Debate on the Exploitation of Natural Plant Diversity to Create Late Blight Resistance in Potato

This paper reports on a debate on intriguing propositions relating to the scientific, agronomic, societal and economic impact of the BIOEXPLOIT project, focusing on late blight resistance in potato. It discusses (i) whether identifying pathogen effectors will facilitate selecting durable R genes, (ii) whether breeding for durable late blight resistance requires deploying Rpi (for Resistance to P hytophthora i nfestans) genes, (iii) whether breeding strategies and cultural practices determine the durability of new resistance genes, (iv) whether marker-assisted breeding for Phytophthora infestans resistance is already in the stage of adoption, (v) to what extent genetically-modified organism technology can advance realizing late-blight resistant potato cultivars, and (vi) whether modifying R genes will result in novel broad spectrum resistance.     

Cisgenesis Does Not Solve the Late Blight Problem of Organic Potato Production: Alternative Breeding Strategies

Average potato yields in Dutch organic farming systems vary from 15 to 29 Mg/ha and are limited by low input of nitrogen and severe late blight attacks caused by the oomycete Phytophthora infestans. Under Dutch late blight regulations it is mandatory to kill the haulm at 7% infestation. The late blight attacks have been so early in the organic potato production of the past few years that its acreage is now gradually decreasing whereas consumer demand is increasing. Agronomic control strategies have limited success. First priority lies in breeding for highly resistant varieties to safeguard organic potato production in the Netherlands. Cisgenesis, however, is not an option for the organic sector. Although the product of cisgenesis does not contain genes from non-crossable species it is a result of a genetic engineering process which is excluded from use in organic agriculture. As the principles and standards of organic agriculture are process-based, cisgenesis does not comply with the norms and standards of organic agriculture. The arguments of the organic sector go well beyond the alleged risks of the gene technology and relate to the technology itself. Breeding at DNA-level, instead of at whole-plant level, violates the integrity of life as described in the concept of naturalness. The Dutch organic sector is now aiming at increasing the traditional breeding activities including the participation of farmer-breeders in close cooperation with the formal breeding companies. Additional selection methods need to be developed to include required traits other than late blight resistance, such as nutrient efficiency. Recently two varieties have been released with high resistance against late blight based on introgressing genes from Solanum bulbocastanum. Organic agriculture can benefit from marker assisted breeding to achieve adequate pyramiding of different, new sources of resistance.     

High-Resolution Mapping of Two Broad-Spectrum Late Blight Resistance Genes from Two Wild Species of the Solanum circaeifolium Group

High levels of resistance to Phytophthora infestans in Solanum are predominantly based on the gene-for-gene interaction. Identification of hitherto unknown R genes is essential for future pyramiding approaches. This could be achieved either through classic introgression breeding or through cisgenesis and could lead to sustainable control of late blight. Here, we report on the mapping of Rpi-cap1 and Rpi-qum1, two late blight R genes identified in the wild species Solanum capsicibaccatum and Solanum circaeifolium ssp. quimense, respectively, to very similar positions on the long arm of chromosome 11. Despite the difficulties encountered for marker development, a high-resolution genetic map with cleaved amplified polymorphic sequence markers was constructed. Furthermore, an R gene cluster-directed profiling approach led to the development of markers that closely linked to or co-segregated with the Rpi-cap1 gene. Both R genes are hypothesized to be homologous to the N gene, a toll-interleukin1 receptor–nucleotide-binding site–leucine-rich repeat domain type of R gene to tobacco mosaic virus from tobacco. To confirm this hypothesis, cloning of Rpi-cap1 and Rpi-qum1 should be pursued. Cloning would also be instrumental to facilitate the introduction of these valuable R genes into potato crops using cisgenic- and marker-assisted breeding approaches.     

Progress in Mapping and Cloning Qualitative and Quantitative Resistance Against <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in Potato and Its Wild Relatives

Cultivated potato is susceptible to many pests and pathogens, none of which is more of a threat to potato agriculture than the late blight disease, caused by the oomycete Phytophthora infestans (Mont.) de Bary. To date all efforts to thwart this most adaptive of pathogens have failed, and early attempts to deploy ‘R genes’ introgressed from the wild Mexican hexaploid Solanum demissum ended in abject failure. With the advent of facile gene mapping and cloning, allied to knowledge of plant resistance gene structure, renewed efforts are leading to mapping and isolation of new sources of late blight resistance in potato wild species, many of which are being performed under the auspices of the BIOEXPLOIT project (Sub-project 2). We document recent advances in late blight resistance gene mapping and isolation, and postulate how these genes, allied to knowledge of pathogen effectors and their recognition specificity, may greatly enhance our chances of halting the progress of late blight disease in potato crops worldwide.     

Resistance to <Emphasis Type="Italic">Potato virus Y</Emphasis> in a Multitrait Potato Breeding Scheme without Direct Selection in Each Generation

In an experimental breeding scheme to improve late blight (Phytophthora infestans) and white potato cyst nematode (Globodera pallida) resistance of tetraploid potato over three generations of crossing and selection, 15 clones survived the final selection, and these were derived from 15 great-grandparents. There was no direct selection for resistance to Potato virus Y (PVY), but 14 out of the 15 great-grandparents were resistant to PVY and three had extreme resistance. Thirteen of the 15 descendants had PVY resistance and one extreme resistance. This was within the range expected for a random (unselected) sample from the genotypes of the great-grandparents. Hence, we found no evidence for any positive or negative association between PVY resistance and the attributes selected. The conclusion is that laborious selection is not required in every generation when many parents have PVY resistance, including some with more than one copy of a PVY resistance gene or resistance at more than one locus. However, in the future, determining the major virus resistance genes present in potential parents in each generation using diagnostic molecular markers would prevent susceptible × susceptible crosses being made and maximise the number of resistant × resistant ones.     

Societal Costs of Late Blight in Potato and Prospects of Durable Resistance Through Cisgenic Modification

In the European Union almost 6 Mha of potatoes are grown representing a value of close to €6,000,000,000. Late blight caused by Phytophthora infestans causes annual losses (costs of control and damage) estimated at more than €1,000,000,000. Chemical control is under pressure as late blight becomes increasingly aggressive and there is societal resistance against the use of environmentally unfriendly chemicals. Breeding programmes have not been able to markedly increase the level of resistance of current potato varieties. New scientific approaches may yield genetically modified marker-free potato varieties (either trans- and/or cisgenic, the latter signifying the use of indigenous resistance genes) as improved variants of currently used varieties showing far greater levels of resistance. There are strong scientific investments needed to develop such improved varieties but these varieties will have great economic and environmental impact. Here we present an approach, based on (cisgenic) resistance genes that will enhance the impact. It consists of five themes: the detection of R-genes in the wild potato gene pool and their function related to the various aspects in the infection route and reproduction of the late blight causing pathogen; cloning of natural R-genes and transforming cassettes of single or multiple (cisgenic) R-genes into existing varieties with proven adaptation to improve their value for consumers; selection of true to the wild type and resistant genotypes with similar qualities as the original variety; spatial and temporal resistance management research of late blight of the cisgenic genetically modified (GM) varieties that contain different cassettes of R-genes to avoid breaking of resistance and reduce build-up of epidemics; communication and interaction with all relevant stakeholders in society and transparency in what research is doing. One of the main challenges is to explain the different nature and possible biological improvement and legislative repercussions of cisgenic GM-crops in comparison with transgenic GM-crops. It is important to realize that the present EU Directive 2001/18/EC on GM crops does not make a difference between trans- and cisgenes. These rules were developed when only transgenic GM plants were around. We present a case arguing for an updating and refinement of these rules in order to place cisgenic GM-crops in another class of GM-plants as has been done in the past with (induced) mutation breeding and the use of protoplast fusion between crossable species.     

Applied Biotechnology to Combat Late Blight in Potato Caused by <Emphasis Type="Italic">Phytophthora Infestans</Emphasis>

Potato is an important crop, grown worldwide. It suffers from many pests and diseases among which late blight, caused by the oomycete Phytophthora infestans, is the worst. The disease is still causing major damage in many potato production areas and control is only possible by applying fungicides frequently. The knowledge on the molecular biology and genetics of the interaction between the plant and the oomycete is developing rapidly. These are relevant fields of study, currently dominated by the discovery of many resistance genes and numerous effector proteins and the analysis of their specific mode of action. These studies may yield essential information needed for the development of durable resistance. The long-term and worldwide effort to breed for resistance so far has had little effect. A novel breeding approach may change this. It is based on cisgenic modification (CM) consisting of marker-free pyramiding of several resistance genes and their spatial and temporal deployment yielding dynamic varieties that contain potato genes only. It is envisioned that this CM approach with potato’s own genes will not only prove societally acceptable but may also result in simplifications in the legislation on use of the CM approach. Various parties in the potato research arena intend to cooperate in this novel approach in a number of developing countries where potato substantially contributes to food security. The use of resources such as land, water and energy improves when the effect of late blight is markedly reduced.     

Cisgenesis,a New Tool for Traditional Plant Breeding,Should be Exempted from the Regulation on Genetically Modified Organisms in a Step by Step Approach

Modern potato breeding requires over 100,000 seedlings per new variety. Main reasons are (1) the increasing number of traits that have to be combined in this tetraploid vegetatively propagated crop, and (2) an increasing number of traits (e.g., resistance to biotic stress) originates from wild species. Pre-breeding by introgression or induced translocation is an expensive way of transferring single traits (such as R-genes, coding for resistance to biotic stress) to the cultivated plant. The most important obstacle is simultaneous transfer of undesired neighbouring alien alleles as linkage drag. Stacking several genes from different wild sources is increasing this linkage drag problem tremendously. Biotechnology has enabled transformation of alien genes into the plant. Initially, transgenes were originating mainly from microorganisms, viruses or non-crossable plant species, or they were chimeric. Moreover, selection markers coding for antibiotic resistance or herbicide resistance were needed. Transgenes are a new gene source for plant breeding and, therefore, additional regulations like the EU Directive 2001/18/EC were developed. Because of a strong opposition against genetic modification of plants in Europe, the application of this Directive is strict, very expensive, hampering the introduction of genetically modified (GM) crops and the use of this technology by small and medium-sized enterprises (SMEs). Currently, GM crops are almost the exclusive domain of multinationals. Meanwhile, not only transgenes but also natural genes from the plant species itself or from crossable plant species, called cisgenes, are available and the alien selection genes can be avoided in the end product. This opens the way for cisgenic crops without alien genes. The existing EU directive for GM organisms is not designed for this new development. The cisgenes belong to the existing breeders’ gene pool. The use of this classical gene pool has been regulated already in agreements regarding breeders’ rights. We are proposing a step by step approach starting with a crop and gene specific derogation and monitoring towards a general exemption of cisgenic plants from the Directive. Two examples, i.e. development of cisgenic potato for resistance to Phytophthora infestans and cisgenic apple for resistance to Venturia inaequalis are discussed shortly for illustration of the importance of cisgenesis as a new tool for traditional plant breeding. Cisgenesis is simplifying introgression and induced translocation breeding tremendously and is highly recommended for SMEs and developing countries.     

小麦地方品种黄方柱中赤霉病主效抗性位点 Fhb1的精细定位

位于小麦3BS染色体上的赤霉病抗性主效基因Fhb1的效应较大而且比较稳定,是培育抗赤霉病品种最常用和最可靠的位点。我国小麦地方品种黄方柱（HFZ）高抗赤霉病且在3BS染色体上携带Fhb1基因。本研究对HFZ/Wheaton重组自交系（RIL）群体的Fhb1位点进行分子标记检测,筛选到两个杂合子株系,并对这两个杂合子株系连续自交3代,衍生出2个次级分离群体（L30和L112）,将这两个次级分离群体在扬州大学农学院试验田种植,并在扬花期进行单花滴注接种鉴定赤霉病,调查这两个群体中各株系的病小穗数、病小穗率、病粒数和病粒率。结果表明,L30和L112两个群体的病小穗数和病小穗率均显著正相关,病粒数和病粒率均显著正相关。同时依据小麦3B染色体参考序列开发了7个新的多态性标记,分别是 X1550、X2214、X2300、X2357、X2455、X2460和X2467。用这些多态性分子标记进行基因型检测,在目标位点发现5个交换株系,根据基因型和病小穗率,进一步将Fhb1定位在分子标记 X2214与 X2357之间,在ctg0954b上这两个分子标记之间的物理距离约为143 kb,NCBI预测该区间有3个候选基因,但是FGENESH在相同区段预测了29个基因,说明目标区段所具有的基因数目很可能多于3个,但少于29个。     

小麦抗条锈基因Yr1和Yr2分子检测体系研究

为建立小麦抗条锈病基因分子检测体系,分别以Yr1和Yr2紧密连锁的标记为检测标记,以Sull-1、CYR29、CYR32、CYR33和V26/CM42为小麦条锈菌鉴别菌株,构建Yr1和Yr2分子检测体系,并对181份小麦高代系材料进行了抗病鉴定和Yr1和Yr2基因分析。结果表明,gwm372和gwm382可作为Yr1的检测标记;wmc364可作为Yr2的检测标记;5个鉴别菌株对Yr1和Yr2具有较好的区分能力。Yr1和Yr2基因在181份小麦高代系材料中比例较低,表明这两个基因在我国小麦抗病育种过程中丢失严重。     

Genetic Improvement of Rice for Bacterial Blight Resistance: Present Status and Future Prospects

The production and productivity of rice has been challenged due to biotic and abiotic factors. Bacterial blight (BB) disease, caused by Xanthomonas oryzae pv. oryzae, is one of the important biotic stress factors, which reduces rice production by 20%–50%. The deployment of host plant resistance is the most preferred strategy for management of BB disease, and breeding disease resistant varieties remains a very economical and effective option. However, it is difficult to develop rice varieties with durable broad-spectrum resistance against BB using conventional approaches alone. Modern biotechnological tools, particularly the deployment of molecular markers, have facilitated the cloning, characterization and introgression of BB resistance genes into elite varieties. At least 46 BB resistance genes have been identified and mapped from diverse sources till date. Among these, 11 genes have been cloned and characterized. Marker-assisted breeding remains the most efficient approach to improve BB resistance by introducing two or more resistance genes into target varieties. Among the identified genes, xa5, xa13 and Xa21 are being widely used in marker-assisted breeding and more than 70 rice varieties or hybrid rice parental lines have been improved for their BB resistance alone or in combination with genes/QTLs conferring tolerance to other stress. We review the developments related to identification and utilization of various resistance genes to develop BB resistant rice varieties through marker-assisted breeding.     

Inheritance of resistance to late blight in foliage and tubers of progenies from parents differing in resistance

Summary TenSolanum tuberosum genotypes differing in resistance to late blight (Phytophthora infestans (Mont.) de Bary) in foliage and tubers were intercrossed to determine the inheritance of disease resistance in their progenies. Plots of 10–15 clones per progeny were established in each of 2 years and resistance assessed by field or laboratory tests. The parental genotypes were similarly tested each year. The parents differed in general combining ability (GCA) for both foliage blight (FB) and tuber blight (TB). The parental and GCA scores were significantly correlated for both aspects of the disease, but the correlations between foliage and tuber scores for parents and for GCAs were not significant. Three parental genotypes were highly resistant in both foliage and tubers, and the genotype with the highest GCA for resistance to both FB and TB (cv. stirling) is recommended as the best parent. There was no evidence of strong genetic correlation between both aspects of resistance, and it is suggested that both be selected for in a breeding programme.     

小麦粒重主效QTL近等基因系的构建和效应评价

粒重是影响小麦产量的主要因素之一。QGw.nau-5A是一个从我国小麦骨干亲本南大2419中鉴定的粒重主效QTL。为评价该QTL不同等位基因对粒重的效应及在育种中的应用潜力,利用分子标记辅助选择技术,分别将南大2419和早洋麦的QGw.nau-5A区段导入望水白和川麦42,构建了不同背景的近等基因系,并比较了不同背景下粒重QTL的效应。结果表明,QGw.nau-5A能在不同背景下显著提高小麦粒重,与轮回亲本相比,近等基因系的百粒重显著增加0.2~0.6g。QGw.nau-5A等位变异对粒重的贡献存在差异,与川麦42的等位变异相比,南大2419和早洋麦的等位变异均能增加粒重,但后者效应更大。     

Relative Braunfäuleresistenz beiSolanum tuberosum ssp.andigena

Zusammenfassung Ergebnisse systematischer Prüfung auf relative Resistenz der Knollen von 1274 Herkünften der ssp.andigena (adg) genenüberPhytophthora infestans werden mitgeteilt. Die Methode des Kn?llchenstestes erm?glichte dabei das Eindringen des Pilzes direkt ins Knollenmark oder über die intakte Schale. Nur etwa 1% der Herkünfte enthielt einen Anteil von >25% Idiotypen mit gutem Resistenzniveau. Eine Analyse der Fundorte ergab keine Anhaltspunkte für die gezielte Suche von Resistenzquellen. Ergebnisse aus der züchterischen Nutzung von adg betreffen die ph?notypische Resistenz der Nachkommen und weitere Merkmale.

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Summary Results are reported of late blight resistance inS. tuberosum ssp.andigena (adg) and its use in breeding. Tests were carried out on material of the gene bank in Gross Lüsewitz (GLKS). Seedlings were cultivated under glass cover in a 6-years test period. To examine tuber resistance toPhytophthora infestans the following test was set up: 2–5 tubers per idiotype, up to 8 idiotypes per accession and year of test were taken and 1274 different accessions were examined with an average number of 23 idiotypes and 59 tubers from each. Inoculation was made by fresh deep cuts to the pith and by subsequent immersion in a solution of zoospores, 6–9×10⁴ ml⁻¹ with a mixture of the pathotypes (1.2.3.4.5.7.10.11, 1.2.3.4.7.8.10 and 1.2.3.4) at a temperature of 16–18°C and at an air humidity of >90%. Evaluation of the presence of aerial mycelium and browning after 7 days allowed the calculation of a resistance score (B_K): grade 9 (highly resistant) to grade 1 (highly susceptible). The mean values derived from the accessions (Fig. 1) revealed that a medium level of susceptibility is dominating and that they rarely exceed average levels of cultivars. Approximately 1% of the accessions contained ≥25% of idiotypes with B_K ≥6 and these may be interesting for breeding purposes. Table 1 shows the analysis of accessions with the most favourable mean values. Geographic evaluation did not yield indications for a more purposeful search for sources of resistance, since detailed reports on find-spots were missing (Table 2). Combination tests withPhytophthora-resistant genes of adg for crosses of adg withS. demissum (dms) andS. stoloniferum (sto) proved to be more useful as they partly showed good heritable resistance to haulm and tuber infection ofP. infestans (Table 3). The problem of the possible role of unknown r-genes is discussed. Only a few crosses with ssp.tuberosum (tbr) generated a progeny with sufficient resistance. Selection from such progeny over several years resulted after the second year in rejections, mainly because of susceptibility for viruses, too small tubers and lack of flowering (Table 4). Results of varietal research of the former German Democratic Republic proved that with the use of suitable parents in breeding for nematode resistance,Phytophthora susceptibility from adg could be balanced off (Table 5). The cultivar Auralia is a good example of the importance of adg in achieving a high level of resistance to late blight.

     

Transferring Translucent Endosperm Mutant Gene Wx-mq and Rice Stripe Disease Resistance Gene Stv-b by Marker-Assisted Selection in Rice (Oryza sativa)

A high-yielding japonica rice variety, Wuyunjing 7, bred in Jiangsu Province, China as a female parent was crossed with a Japanese rice variety Kantou 194, which carries a rice stripe disease resistance gene Stv-bⁱ and a translucent endosperm mutant gene Wx-mq. From F₂ generations, a sequence characterized amplified region (SCAR) marker tightly linked with Stv-bⁱ and a cleaved amplified polymorphic sequence (CAPS) marker for Wx-mq were used for marker-assisted selection. Finally, a new japonica rice line, Ning 9108, with excellent agronomic traits was obtained by multi-generational selection on stripe disease resistance and endosperm appearance. The utilization of the markers from genes related to rice quality and disease resistance was helpful not only for establishing a marker-assisted selection system of high-quality and disease resistance for rice but also for providing important intermediate materials and rapid selection method for good quality, disease resistance and high yield in rice breeding.     

基于GBS技术构建四倍体杂交冰草超高密度遗传连锁图谱

冰草是优良的牧草,也是小麦等麦类作物抗逆遗传改良的重要基因库。构建冰草超高密度遗传连锁图谱,有利于冰草的标记辅助育种和遗传分析。本研究利用基因分型测序（GBS）技术,对四倍体杂交冰草F₂分离群体的115个单株及其亲本蒙古冰草和航道冰草进行高通量SNP基因分型,经测序获得了超过584 Gb的数据,SNP鉴定以及完整度和偏分离过滤后,最终得到了5 035个高质量的SNP标记;采用Join Map 4.0作图软件构建四倍体冰草超高密度遗传连锁图谱,该遗传图谱包括14个连锁群,各连锁群的长度范围在68.959～280.309 cM之间,平均长度为153.473 cM,覆盖的基因组总长度为  2 148.621 cM,标记间的平均距离为0.427 cM。该图谱是目前冰草中分子标记数目最多、密度最高的遗传连锁图谱,为进一步开展冰草品质、抗性等重要性状的QTL定位、图位克隆及分子标记辅助育种研究等奠定了基础。     

Correlation between resistance to late blight in foliage and tubers in potato clones from parents of contrasting resistance

Summary The relationship between resistance to late blight in foliage and tuber was examined on 50 clones of each of five progenies from crosses where one parent was resistant in both foliage and tuber and the other susceptible. Foliage resistance was assessed in a field trial and tuber resistance in a laboratory test on glasshouse-grown tubers. The genetical and environmental components of variance and the phenotypic and genetical correlations between foliage and tuber blight scores were determined for each progeny. Foliage and tuber resistance were correlated, indicating either that both are determined by the same genes or by different linked genes. The environmental component of variation was greater for tuber blight than foliage blight. We suggest that the most effective way of selecting for resistance to both aspects of the disease in a breeding programme is to select those resistant in the foliage and then screen them for tuber resistance.     

QTL Analysis of Late Blight Resistance in a Diploid Potato Family of Solanum spegazzinii × S. chacoense

Several quantitative trait loci (QTL) for resistance to late blight have been reported in diploid potatoes. The diploid wild potato species Solanum chacoense possesses a high degree of horizontal resistance to late blight. In the present study, we report on QTL mapping for late blight resistance in a diploid mapping population of 126 F₁ of Solanum spegazzinii (susceptible) × S. chacoense (resistant). The area under the disease progress curve (AUDPC) values for late blight resistance using the “whole plant in vitro assay” and the “detached leaf assay” on the mapping population displayed quantitative variation. Out of 64 AFLP primer-pairs combinations and SSR markers, a total of 209 significant AFLP loci were placed onto the 12 linkage group of potato covering a total map length of 6548.1 cM. QTL analysis based on the AUDPC dataset of the “whole plant in vitro assay” using the interval-mapping option identified two QTL (LOD?>?2.5) located on linkage groups IX and X, which explained 14.70 and 3.40% variation, respectively. The present study revealed the presence of potential new genetic loci in the diploid potato family contributing to quantitative resistance against late blight.     

小麦水分高效利用分子育种研究进展

干旱缺水是引起全球作物减产的主要自然灾害。小麦水分高效利用遗传改良是一个重大课题。本文对小麦水分高效利用分子标记、功能基因克隆鉴定、转基因、分子设计育种等方面的国内外研究进展进行了综述。在小麦中已经鉴定出A组染色体上存在控制水分利用效率的QTL;开发出与TaSAP1等基因紧密连锁的功能标记;克隆了转录因子基因TaWRKY并对其抗旱抗逆功能进行了鉴定;转HVA1和DREB等基因的小麦可以明显提高水分高效利用能力;利用水旱地品种进行轮回杂交结合分子标记辅助选择育种技术进行了小麦水分高效利用分子设计育种初探。以上研究进展为小麦水分高效利用遗传改良提供了理论依据和技术支撑。     

Dissection of gene loci underlying pasting temperature in cassava

In this study, a fine genetic map within the quantitative trait loci (QTL) underlying pasting temperature (PT) of cassava (Manihot esculenta Crantz) was constructed using newly developed simple sequence repeat (SSR) markers. The SSRs were designed on the basis of two scaffolds (S11341 and S4043) of the cassava genome, which covered previously identified QTL regions of the PT trait. A total of 55 and 61 SSR markers derived from S11341 and S4043, respectively, representing 0.29% of the cassava genome, were generated; of which 23 and 19 showed informative polymorphic patterns. Consequently, all identified informative polymorphic markers were used to genotype 200 F₁ progeny plants. The genotypic data were then analyzed, and the results showed that 480 markers were distributed across 23 linkage groups (LGs) with total length of 1,334 centimorgans (cM). An analysis of QTL underlying the PT trait revealed that marker EME81 on LG 1 had significant associations (P < 0.0001) in all environments evaluated. Four candidate genes were identified and selected for gene expression analysis in the parents, and among F₁ lines with high and low PT values. Significant differences were observed in relative expression of carbohydrate phosphorylase (CP) and starch synthase II (SSII) between high and low PT in 6-month-old cassava. We found CP and SSII genes to potentially control the PT trait. In addition, the marker EME81 was found to be a promising marker for specific PT trait selection in cassava populations, which should facilitate marker-assisted selection for desired PT traits.