Identification of QTLs for Starch Content in Sweetpotato （Ipomoea batatas （L.） Lam.）

Sweetpotato （Ipomoea batatas （L.） Lam.） breeding is challenging due to its genetic complexity. In the present study, interval mapping （IM） and multiple quantitative trait locus （QTL） model （MQM） analysis were used to identify QTLs for starch content with a mapping population consisting of 202 F1 individuals of a cross between Xushu 18, a cultivar susceptible to stem nematodes, with high yield and moderate starch, and Xu 781, which is resistant to stem nematodes, has low yield and high starch content. Six QTLs for starch content were mapped on six linkage groups of the Xu 781 map, explaining 9.1-38.8% of the variation. Especially, one of them, DMFN 4, accounted for 38.8% of starch content variation, which is the QTL that explains the highest phenotypic variation detected to date in sweetpotato. All of the six QTLs had a positive effect on the variation of the starch content, which indicated the inheritance derived from the parent Xu 781. Two QTLs for starch content were detected on two linkage groups of the Xushu 18 map, explaining 14.3 and 16.1% of the variation, respectively. They had a negative effect on the variation, indicating the inheritance derived from Xu 781. Seven of eight QTLs were co-localized with a single marker. This is the first report on the development of QTLs co-localized with a single marker in sweetpotato. These QTLs and their co-localized markers may be used in marker-assisted breeding for the starch content of sweetpotato.     

Identification of QTLs for Starch Content in Sweetpotato(Ipomoea batatas(L.) Lam.)

Sweetpotato（Ipomoea batatas（L.）Lam.）breeding is challenging due to its genetic complexity.In the present study,interval mapping（IM）and multiple quantitative trait locus（QTL）model（MQM）analysis were used to identify QTLs for starch content with a mapping population consisting of 202 F1 individuals of a cross between Xushu 18,a cultivar susceptible to stem nematodes,with high yield and moderate starch,and Xu 781,which is resistant to stem nematodes,has low yield and high starch content.Six QTLs for starch content were mapped on six linkage groups of the Xu 781 map,explaining 9.1-38.8%of the variation.Especially,one of them,DMFN₄,accounted for 38.8%of starch content variation,which is the QTL that explains the highest phenotypic variation detected to date in sweetpotato.All of the six QTLs had a positive effect on the variation of the starch content,which indicated the inheritance derived from the parent Xu 781.Two QTLs for starch content were detected on two linkage groups of the Xushu 18 map,explaining 14.3 and 16.1%of the variation,respectively.They had a negative effect on the variation,indicating the inheritance derived from Xu 781.Seven of eight QTLs were co-localized with a single marker.This is the first report on the development of QTLs co-localized with a single marker in sweetpotato.These QTLs and their co-localized markers may be used in marker-assisted breeding for the starch content of sweetpotato.     

低钾胁迫对甘薯块根淀粉理化特性的影响及其基因型差异

【目的】甘薯[Ipomoea batatas(L.)Lam]属块根类淀粉作物,同时也是较典型"喜钾"作物,而甘薯种植区土壤缺钾十分普遍。同时,甘薯块根淀粉用途与其理化特性密切相关,因此,必须加强研究钾缺乏对甘薯块根淀粉理化特性的影响。【方法】选用3个不同钾效率型甘薯徐薯32(耐低钾与钾高效型)、徐薯18(中间型)与宁紫薯1号(不耐低钾与钾低效型)为材料,利用1980年设置的长期肥料定位试验田低钾(LK,氮磷处理)与正常钾水平(CK,氮磷钾处理)两处理,分析甘薯块根淀粉粒径大小与分布、热焓特性、糊化特性、吸湿性及膨胀力等特征指标差异。【结果】与正常钾相比,低钾胁迫改变甘薯块根中直链淀粉、支链淀粉、总磷、总脂肪和氮等主要组分,不同钾效率型甘薯间变化存在差异。甘薯淀粉粒径存在≤2.50μm、2.50—5.00μm和5.00—30.00μm等明显的3峰分布,低钾胁迫下淀粉粒径及其分布均发生变化,淀粉平均粒径降低,其中在体积分布中,徐薯32平均粒径减幅最小,宁紫薯1号降幅达显著水平(P0.05),表面积与数目分布中差异不显著,而不同粒径的百分比、平均粒径与峰值(数目分布中无峰值)均有不同的变化,且这些变化在不同品种间也存在差异。淀粉糊化过程中的起始温度(T_0)、峰值温度(T_p)和热焓值(ΔH)普遍低于正常钾供应处理的相应指标,尤其宁紫薯1号在3个热特征值上的差异均达显著水平,其差异幅度大于徐薯32,3个甘薯品种最高粘度值、崩解值均明显下降,徐薯18与宁紫薯1号的最低粘度值、最终粘度值受低钾影响显著上升,而徐薯32的表现则相反。块根淀粉吸湿性均不同程度降低,徐薯18与宁紫薯1号降低达显著水平(P0.05),徐薯32淀粉吸湿性变化较小,但其吸湿性值最高。淀粉膨胀力不同程度上升,宁紫薯1号增幅达显著水平(P0.05),徐薯32淀粉颗粒膨胀力较高,受低钾胁迫影响效应小。相关分析表明,甘薯淀粉主要理化指标间密切相关,低钾胁迫改变甘薯块根淀粉组分及颗粒粒径大小,是导致不同程度影响甘薯块根淀粉的糊化特性、热特性等理化特性的主要因素。【结论】钾是影响甘薯块根淀粉理化特性的重要元素,低钾胁迫对甘薯块根淀粉组分及其品质理化特性产生较明显影响,不同钾效率型甘薯块根淀粉特性对低钾胁迫响应不同。     

Regulation of Anthocyanin during Storage Root Development Stage in Sweet Potato(Ipomoea batatas(L.) Lam.)

Anthocyanin accumulation during storage root development in purple-fleshed sweet potato was analyzed by detection of anthocyanin concentration, accumulation rate and the expression pattern of anthocyanin biosynthetic genes by semi-quantitative RT-PCR. Anthocyanin concentration in sweet potato cvs Jishu 18 and Ayamurasaki increased steadily during storage root development stage. The accumulation rate in two genotypes peaked at 50 to 65 d after transplanting, and then declined rapidly. During storage root development of Ayamurasaki, the anthocyanin biosynthesis gene, IbCHS, was constitutively expressed, the genes IbF3H, IbDFR, IbANS were induced steadily, reaching a maximum at the later stage of root thickening, and IbPAL steadily decreased. Therefore, the mechanism of anthocyanin accumulation differed between the two cultivars, and anthocyanin biosynthesis was regulated through regulation of its synthetic enzymes.     

不同基因型甘薯（Ｉｐｏｍｏｅａ ｂａｔａｔａｓ Ｌａｍ．）吸收累积Ｃｄ的特征差异性研究

采用旱地小区试验,研究了Cd污染土壤中甘薯吸收累积Cd的品种差异,探讨了甘薯吸收累积Cd的相关机理.结果表明,甘薯品种间不仅生物量差异较大,对Cd的吸收累积能力也不同,8个品种的地上部茎叶生物量(DW)在4 594.9～8 232.2 kg·hm-2,鲜样Cd含量范围为0.032 9～0.057 4 mg·kg-1;地下部块根生物量(DW)在7 809.0～14 269.7 kg·hm-2,鲜样Cd含量范围为0.002 2～0.011 0mg·kg-1,茎叶Cd含量明显高于块根,但均未超过国家食品中污染物限值标准(GB 2762-2005)(根茎类蔬菜<0.1 mg·kg-1).而与无公害蔬菜质量标准(GB 18406.1-2001)进行比较(<0.05 mg·kg-1),有3个甘薯品种(心香、湘薯15、泉薯9号)的茎叶Cd含量超标.因此,轻度Cd污染土壤种植的甘薯其茎叶可能存在摄食健康风险,而块根基本无风险,若将Cd污染土壤改制种植甘薯,必须妥善处理其地卜部茎叶.     

鲜食甘薯栽培技术研究

在普通生产田和林间生产田,从品种、栽植密度和施用钾肥3方面研究了鲜食甘薯栽培技术.结果表明,所用3个甘薯品种(宁薯192、宁紫1号和济薯18)的商品薯产量构成差异较大,宁薯192为薯重型,商品薯比例高;宁紫1号为薯数型,小薯数量多;济薯18为中间型,商品薯产量较高.在普通生产田,栽植密度1 hm2 48 000株、增施225 kg/hm2硫酸钾,可获得最高的商品薯产量.在已成大树的林间生产田,鲜食甘薯的商品薯产量非常低.     

秋甘薯干物质积累与产量形成

[目的]探讨秋甘薯((I)pomoea batatas Lam.)干物质积累和产量形成规律.[方法]以西成薯007和泉薯9号为试验材料,对秋甘薯干物质积累过程及分配规律进行了研究.[结果]随着生长的推进秋甘薯植株鲜重和干重均增加,干物率缓慢上升,T/R比呈先快后慢的下降趋势,徒长较严重;主要的干物质接收器官大中薯在植后42 ～56 d形成,之后随着薯块的生长大中薯率直线上升;泉薯9号的LAI在整个生长内变化较为平稳,栽插42d后西成薯007的有效光合叶面积呈直线上升,到98 d收获时仍大于9.叶干物质分配率在生长期内呈直线下降,到收获时只占总干物质重的10％;柄干物质分配在生长期内保持在10％左右;茎所分配的干物质从28 d的40％逐渐下降到收获时的20％以下,而块根在28 d出现后其干物质分配呈直线上升,到收获时达到总干重的60％.叶片叶绿素含量在生长期内缓慢下降;甘薯正常生长时,叶片的SOD、POD、CAT等酶的协同变化将MDA含量维持在一个较为平稳的水平,以保证植株的正常生长和同化作用的持续进行.块根可溶性糖含量在甘薯植株受到低温胁迫时急剧上升,在后期持续低温时保持在一个较高的水平,以维持细胞渗透压,最大限度地保证植株不受冷害;块根淀粉含量在栽后28 d形成后即直线上升.在秋甘薯生长的中后期,决根的GA、ABA和IAA含量均较叶片低,植株体内的GA含量栽插56 d后随着植株的衰老逐渐降低.在秋甘薯生长后期ABA含量迅速增加,进一步加快了植株衰老,而LAA含量在这一时期迅速增加应是延缓植株衰老.[结论]该研究对一季薯干倍增技术的研究具有指导意义.     

Accumulation and Gene Expression of Anthocyanin in Storage Roots of Purple- Freshed Sweet Potato [Ipomoea batatas （L.） Lam] Under Weak Light Conditions

Anthocyanidin in plants, an important pigment, is of great interest to researchers, consumers, and commercial entities due to its physiological functions. Anthocyanin content and mRNA levels of anthocynin biosynthesis genes were investigated in storage root of different purple-fleshed sweet potatoes （PFSP） genotypes to understand the regulation mechanism of anthocyanin under weak light conditions. Anthocyanin content, its amount of accumulation, and the expression of CHS, DFR, F3H, GT, and ANS genes in the PFSP storage root under weak light conditions were studied. The results demonstrated that the anthocyanin content of the treatments was decreased and was obviously lower than that of the control until 30 days after shading in Ayamurasaki, while it was lower than that of the control from the beginning of shading in Jishu 18. Their accumulation rates of both treatmeants were lower than its control before 10-20 d of shading in Jishu 18, while those of Ayamurasaki weren＇t in their treatments. This indicated that Jishul 8 is more sensitive to light as compared to Ayamuraska. Under the different weak light conditions, mRNA levels for ibCHS, ibF3H, ibDFR, and ibANS were obviously decreased, while the expression of ibGT was increased. These results indicated that anthocyanin content was regulated by light at the mRNA levels and the enzymatic level in sweet potato. Therefore, the development dynamic response to anthocyanin content varied in different genotypes of PFSP, and mRNA levels of anthocyanin biosynthesis were inhibited under the weak light condition.     

甘薯特异材料BB30-224的特性鉴定与利用

甘薯种质材料BB30-224是从国际马铃薯中心引进的BB系列杂交种子中选育出的品系。经鉴定:BB30-224具有生产力强、干物质含量高、抗黑斑病、贮藏性好、嫁接诱导开花多、结实率高等优良特性,而且还具有干物质含量和黑斑病抗性遗传能力强、后代品系入选率高等特点。BB30-224是现有育种材料中表现较好,优点突出的优秀种质资源。     

SSR fingerprinting of 203 sweetpotato (Ipomoea batatas (L.) Lam.) varieties

Simple sequence repeat (SSR) markers have been shown to be a powerful tool for varieties identification in plants. However, SSR fingerprinting of sweetpotato varieties has been a little reported. In this study, a total of 1294 SSR primer pairs, including 1215 genomic-SSR and 79 expressed sequence tag (EST)-SSR primer pairs, were screened with sweetpotato varieties Zhengshu 20 and Luoxushu 8 and their 2 F₁ individuals randomly sampled, and 273 and 38 of them generated polymorphic bands, respectively. Four genomic-SSR and 3 EST-SSR primer pairs, which showed good polymorphism, were selected to amplify 203 sweetpotato varieties and gave a total of 172 bands, 85 (49.42%) of which were polymorphic. All of the 203 sweetpotato varieties showed unique fingerprint patterns, indicating the utility of SSR markers in variety identification of this crop. Polymorphism information content (PIC) ranged from 0.5824 to 0.9322 with an average of 0.8176. SSR-based genetic distances varied from 0.0118 to 0.6353 with an average of 0.3100 among these varieties. Thus, these sweetpotato varieties exhibited high levels of genetic similarity and had distinct fingerprint profiles. The SSR fingerprints of the 203 sweetpotato varieties have been successfully constructed. The highly polymorphic SSR primer pairs developed in this study have the potential to be used as core primer pairs for variety identification, genetic diversity assessment and linkage map construction in sweetpotato and other plants.     

基于EST-SSR标记甘薯连锁图谱构建及淀粉性状的QTL定位

【目的】利用分子标记技术,构建甘薯遗传连锁图谱,并分析甘薯淀粉含量性状的QTL位点,为高淀粉含量甘薯种质资源利用及甘薯分子标记辅助育种提供理论和实践依据。【方法】以高淀粉含量品种万薯5号为母本、低淀粉含量品种商丘52-7为父本建立杂交群体,利用EST-SSR标记,采用"双假测交"策略和运用Join Map4.0软件,分别构建双亲遗传连锁图谱,并结合F1(2012、2013年)群体表型数据采用区间作图法对淀粉含量性状进行QTL检测。【结果】利用1 679对EST-SSR引物筛选出的1 045对多态性引物检测F1群体的标记基因型,获得了1 418个标记位点。分别对上述获得的父母本多态性标记进行遗传连锁分析,在LOD≥5.0情况下,分别构建父母本的连锁遗传图谱。采用642个标记的多态性位点构建母本连锁群74个,其中,215个标记位点位于连锁图谱上,占标记多态性位点总数的33.5%。每个连锁群上有2—11个标记位点,连锁群长度在0.6—129.4 cM,图谱总长为3 826.07 c M,标记间平均距离为17.80 c M。属于父本的776个标记位点构建了80个连锁群,共有252个标记位点构建在连锁图谱上,占标记总数的32.5%,每个连锁群上有2—24个标记位点,连锁群长度在2.0—156.8 c M,图谱总长为3 955.0 cM,标记间平均距离为15.7 c M。以F1杂交群体构建的遗传连锁图谱,结合2012年、2013年2个环境,利用QTL作图软件MapQTL5.0,采用区间作图法进行分析,共检测到17个与淀粉含量性状相关的QTL,贡献率在8.4%—40.5%。其中qWsc-1、qWsc-2、qWsc-3 3个QTL位于母本万薯5号连锁群上,且在2年环境中均可检测到;14个QTL位于父本商丘52-7连锁群上,qSsc-1、qSsc-2、qSsc-3、qSsc-4、qSsc-8、qSsc-10、qSsc-11、qSsc-12是在2个环境均检测到的QTL。qSsc-5、qSsc-6、qSsc-7、qSsc-9、qSsc-13、qSsc-14是只在1个环境检测到的QTL。标记GDAAS0603在双亲中和2个环境中均同时检测到,这些环境稳定QTL可用于分子标记辅助选择。【结论】分别构建了亲本EST-SSR分子标记连锁群图谱,丰富了构建甘薯图谱的标记类型,定位了17个与淀粉含量相关的QTL位点。     

干旱条件下不同耐旱性甘薯苗期根系抗氧化特性分析（英文）

[目的]揭示不同耐旱性甘薯苗期根系的抗氧化特性。[方法]以耐旱性强的济薯21和耐旱性弱的济紫薯1为材料,采用水培和PEG-6000模拟干旱胁迫的方法 ,研究了干旱条件下不同耐旱性甘薯苗期根系活性氧、抗氧化物质和抗氧化酶差异。[结果]在干旱条件下,济紫薯1苗期根系的H_2O_2含量和O~(2-)生成速率分别比济薯21的高1.9%和102.6%,济薯21和济紫薯1根系中的Vc含量分别增加了25.3%和81.1%,而多酚含量则增加了24.8%和37.6%,在上述指标中济紫薯1的增幅均高于济薯21;干旱条件下,济薯21和济紫薯1根系中的抗氧化酶活性均升高,其中POD活性增幅最大,PPO活性增幅最小,济薯21苗期根系中的SOD、POD、APX和PPO活性增幅比济紫薯1的高一倍左右,两个品种干旱处理间的SOD3、POD2、CAT1、APX2和PPO2-等同工酶的条带变化明显。[结论]耐旱性越强的甘薯品种,干旱条件下苗期根系中H_2O_2含量、O~(2-)-生成速率、Vc和多酚含量增幅越小,抗氧化酶活性增幅越高,抗氧化酶差异主要与SOD3、POD2、CAT1、APX2和PPO2-等同工酶的变化有关。     

Development of SRAP Markers Linked to a Gene for Stem Nematode Resistance in Sweetpotato, Ipomoea batatas (L.) Lam.

Sequence-related amplification polymorphism (SRAP) markers closely linked to stem nematode resistance gene were developed in sweetpotato, Ipomoea batatas (L.) Lam. Using bulked segregant analysis (BSA), 200 SRAP primer combinations were screened with the resistant and susceptible bulked DNA from the 196 progenies of an F1 single-cross population of resistant parent Xu 781×susceptible parent Xushu 18, 77 of them showed polymorphic bands between resistant and susceptible DNA. Primer combinations detecting polymorphism between the two bulks were used to screen both parents and 10 individuals from each of the bulks. The results showed that primer combination A9B4 produced 3 specific bands in the resistant plants but not in the susceptible plants, suggesting that the markers, named Nsp1, Nsp2 and Nsp3, respectively, linked to a gene for stem nematode resistance. Primer combination A3B6 also produced a SRAP marker named Nsp4 linking to the resistance gene. Amplified analysis of the 196 F1 individuals indicated that the genetic distance between these markers and the resistance gene was 4.7, 4.7, 6.3, and 9.6 cM, respectively.     

干旱条件下不同耐旱性甘薯苗期根系抗氧化特性分析

[目的]揭示不同耐旱性甘薯苗期根系的抗氧化特性.[方法]以耐早性强的济薯21和耐旱性弱的济紫薯1为材料,采用水培和PEG-6000模拟干旱胁迫的方法,研究了干旱条件下不同耐旱性甘薯苗期根系活性氧、抗氧化物质和抗氧化酶差异.[结果]在干旱条件下,济紫薯1苗期根系的H2O2含量和O2-生成速率分别比济薯21的高1.9％和102.6％,济薯21和济紫薯1根系中的Vc含量分别增加了25.3％和81.1％,而多酚含量则增加了24.8％和37.6％,在上述指标中济紫薯1的增幅均高于济薯21;干旱条件下,济薯21和济紫薯1根系中的抗氧化酶活性均升高,其中POD活性增幅最大,PPO活性增幅最小,济薯21苗期根系中的SOD、POD、APX和PPO活性增幅比济紫薯1的高一倍左右,两个品种干旱处理间的SOD3、POD2、CAT1、APX2和PPO2等同工酶的条带变化明显.[结论]耐旱性越强的甘薯品种,干旱条件下苗期根系中H2O2含量、O2-生成速率、Vc和多酚含量增幅越小,抗氧化酶活性增幅越高,抗氧化酶差异主要与SOD3、POD2、C AT1、APX2和PPO2等同工酶的变化有关.     

甘薯体细胞胚胎发生遗传变异的RAPD分析

以“郑红５”等６个甘薯品种为材料,通过离体培养诱导体细胞胚胎发生与植株再生。利用ＲＡＰＤ分子标记方法,在ＤＮＡ水平上分析甘薯品种间遗传差异以及体细胞胚胎无性系的遗传变异。筛选了４５个随机引物,其中７个引物ＰＣＲ扩增后在品种间和体细胞胚胎无性系中表现ＲＡＰＤ多态性,并在相应胚性愈伤组织中检测到少数ＲＡＰＤ变异位点。证明利用ＲＡＰＤ分子标记方法鉴定甘薯体细胞胚胎发生遗传变异的可行性,以及为通过离体培养     

超声波提取紫薯多糖的工艺优化

[目的]优化紫薯多糖的提取工艺。[方法]利用超声波技术提取紫薯粗多糖,用单因素试验和正交试验设计相结合的方法获得最佳提取工艺。[结果]影响紫薯多糖得率的主要因素按重要性排序为：超声频率〉料液比〉提取温度〉提取时间;紫薯粗多糖最佳提取工艺条件为：料液比1∶15,提取时间40 min,提取温度60℃,超声频率80 kHz,此条件下紫薯多糖的提取率为3.63%。[结论]利用超声技术提取紫薯多糖可以提高多糖得率、缩短提取时间、采用较低温度提取、节省提取溶剂,降低提取成本。     

紫心甘薯花青素积累与PAL活性的关系

以块根着色程度不同的2个紫心甘薯品系‘A3’和‘山川紫’为试材,以白心甘薯‘禺北白’为对照,对其不同器官(叶、茎和块根)的花青素含量和苯丙氨酸解氨酶(PAL)的活性进行了测定,对它们之间的变化趋势进行了相关性分析并建立了回归模型.结果表明:不论是不同品系的整株水平,还是同一品系的不同器官,着色程度深的部位,花青素的含量较高,PAL活性也较高.相关性分析以及回归模型显示,花青素含量与PAL活性之间呈极显著的线性正相关.说明PAL是紫心甘薯花青素合成的关键酶.     

紫心甘薯花青素积累与PAL活性的关系

以块根着色程度不同的2个紫心甘薯品系‘A3'和‘山川紫'为试材,以白心甘薯‘禺北白'为对照,对其不同器官(叶、茎和块根)的花青素含量和苯丙氨酸解氨酶(PAL)的活性进行了测定,对它们之间的变化趋势进行了相关性分析并建立了回归模型.结果表明:不论是不同品系的整株水平,还是同一品系的不同器官,着色程度深的部位,花青素的含量较高,PAL活性也较高.相关性分析以及回归模型显示,花青素含量与PAL活性之间呈极显著的线性正相关.说明PAL是紫心甘薯花青素合成的关键酶.

Abstract：

The contents of anthocyanin and the enzyme activities of phenylalanine ammonialyase(PAL)in different organs of three strains of sweet potato[Ipomoea batatas(L.)Lam.],including two purple-fleshed strains(A3 and Yamakawamurasaki)and one white-fleshed strain(Yubeibai)were determined.The relationship between anthocyanin content and PAL activity was investigated by correlation analysis,and a linear regression model was established.The results indicated that the activity of PAL was significantly and positively correlated with the content of anthocyanin,whether in different organs of the same strain or in the whole plant of different strains,thus indicating that PAL is one of the key enzymes in anthocyanin synthesis in purple-fleshed sweet potato.     

超声波辅助提取甘薯根颈多糖

甘薯根颈是甘薯生产中的大宗副产物,是提取功能性多糖的廉价资源。通过正交试验,对甘薯根颈多糖超声波辅助提取的影响因素进行了筛选和优化。结果表明：料液比、超声波频率、浸提温度、超声波功率、超声浸提时间等因素对提取率影响较大,较为合理的提取方案为：料液比1：30,超声波频率26kHz,超声波功率350W,浸提温度60℃,浸提时间30min。以此优化方案提取3次,提取物的平均提取得率为4.95%。通过棕色环试验、蒽酮-硫酸法、Fehling试剂法、双缩脲法证实提取物为多糖。