籼型杂交稻淀粉RVA谱特征值的杂种优势分析

以6个籼型三系不育系和5个籼型恢复系按不完全双列杂交设计配制的30个杂交稻组合及其亲本为材料,对其籽粒淀粉RVA谱各特征值进行了测定和分析。结果表明：（1）亲本间和杂交稻组合间的淀粉RVA谱各特征值均存在极显著的差异,其中变异最大的是消减值,最小的是糊化开始温度。（2）杂交稻组合淀粉RVA谱各特征值的变异系数均小于杂交稻亲本的变异系数;峰值粘度、崩解值、最低粘度和最终粘度的平均值杂交稻组合大于亲本,其他性状的平均值杂交稻组合小于亲本。（3）峰值粘度存在极显著的正向超亲优势;崩解值、最低粘度、最终粘度和回复值存在极显著的正向中亲优势;消减值和峰值时间存在极显著的负向中亲优势和正向低亲优势;糊化开始温度存在极显著的负向低亲优势;最终粘度、回复值和糊化开始温度表现为极显著的负向竞争优势,其他性状表现为正向竞争优势。（4）峰值粘度、崩解值、消减值和回复值等4个性状,各杂交稻组合与其母本、中亲值呈现极显著正相关,峰值时间与中亲值呈显著正相关。而与其父本的相关均未达显著水平。     

不同类型土壤上种植的小麦籽粒中与淀粉合成相关的酶活性变化

测定池栽条件下灰潮土、水稻土、砂姜黑土上种植的强筋小麦‘郑麦9023’籽粒灌浆过程中腺苷二磷酸葡萄糖焦磷酸化酶（AGPP）、尿苷二磷酸葡萄糖焦磷酸化酶（UGPP）、可溶性淀粉合成酶（SSS）、淀粉粒结合淀粉合成酶（GBSS）、淀粉分支酶（SBE）5个与淀粉合成有关的酶活性变化的结果表明,不同类型土壤上种植的小麦籽粒中AGPP、UGPP、SSS、GBSS、SBE活性均呈单峰曲线变化,花后18d,AGPP、UGPP、SSS和SBE活性达到峰值,而GBSS则在花后24d达到峰值。AGPP、SSS、SBE活性峰值表现为灰潮土〉水稻土〉砂姜黑土,UGPP峰值表现为灰潮土〉砂姜黑土〉水稻土,GBSS峰值则表现为水稻土〉灰潮土〉砂姜黑土。     

转反义Wx基因水稻颖果中与淀粉合成和积累相关酶活性的变化

将反义Wx基因转入水稻,导致Wx蛋白不同程度减少,颖果中的直链淀粉含量不同程度下降,总淀粉含量显著降低,直链淀粉与总淀粉的比值极显著降低。在水稻颖果发育过程中,ADPG-PPase、GBSS、SSS和SBE的活性在灌浆前期迅速升高,达最大值后很快下降,在灌浆中后期下降趋缓。Wx蛋白减少后的转基因水稻颖果中的GBSS活性明显下降,下降幅度与直链淀粉含量相一致,而且活性高峰期比其亲本有所提前。转基因水稻颖果中ADPG-PPase和SSS的活性在颖果发育的前中期,SBE则在中后期高于相应的亲本。     

花后高温对不同耐热性小麦品种籽粒淀粉形成的影响

以耐热性不同的2个小麦品种济麦20和鲁麦21为材料,分别于花后5～9d（T1）和15～19d（T2）进行高温处理,研究了小麦花后不同阶段高温对籽粒淀粉积累、淀粉粒分布及相关酶活性的影响。结果表明,花后高温显著降低籽粒淀粉积累量;显著降低籽粒淀粉及支链淀粉含量,但提高直链淀粉含量、直／支链淀粉比例。处理间比较,他处理对籽粒淀粉积累的影响程度较T1处理大。品种间比较,高温对济麦20的影响程度较鲁麦21大。高温使A型淀粉粒的体积、数量和表面积比例显著增加,B型淀粉粒的体积、数量和表面积比例显著降低。T1处理后,两品种籽粒蔗糖含量、蔗糖合酶（SS）和腺苷二磷酸葡萄糖焦磷酸化酶（AGPP）、可溶性淀粉合酶（SSS）、束缚态淀粉合酶（GBSS）和淀粉分支酶（SBE）活性均略高于对照;但济麦20、鲁麦21上述指标分别于花后15、20d开始低于对照。他处理后,两品种籽粒蔗糖含量、SS、AGPP、SSS、GBSS和SBE活性显著低于对照,济麦20上述指标的降幅较鲁麦21大。与其它淀粉合成相关酶相比,高温对籽粒GBSS活性的影响程度较小。两品种处理间籽粒蔗糖含量、SS、AGPP、SSS、GBSS及SBE活性的变化趋势,与淀粉积累量的变化趋势基本一致。说明灌浆期高温使籽粒淀粉积累量降低,一方面是由于籽粒蔗糖供应较低引起糖源不足;另一方面则是由于灌浆中后期淀粉合成相关酶活性下降使淀粉合成受抑所致。     

杂交水稻直链淀粉含量遗传分析

杂交水稻稻谷是从Ｆ１代杂合体收获的Ｆ２代种子,由于Ｆ２代属分离群体,直链淀粉含量等稻米品质会发生分离现象．本研究选择直链淀粉含量低、中、高的５个亲本配组的５个杂交组合,按单粒分析法分析双亲亲本、Ｆ１和Ｆ２代的种子的直链淀粉含量．结果表明,Ｆ２代呈现１（低）：３（中＋高）孟德尔分离规律．在５个组合的单粒分析中发现分离的Ｆ２代群体呈双峰的连续分布,该现象表明高直链淀粉含量对低直链淀粉含量为不完全显性,除单一基因控制外,还受微效多基因修饰．因此杂交水稻育种工作中,尤其是优质杂交稻新组合选育时,必须注意选择双亲的直链淀粉含量基本一致,以防分离现象影响米质．     

施氮量和花后土壤含水量对强筋小麦籽粒淀粉合成及品质的影响

以强筋小麦品种'济麦20'为材料,在防雨池栽培条件下研究了施氮量和花后土壤含水量对强筋小麦籽粒淀粉合成及其品质的影响,以明确强筋小麦获得高产优质的花后适宜土壤含水量及施氮量.结果表明:在同一施氮量下,适宜的花后土壤含水量(60%～70%)籽粒游离态淀粉合成酶(SSS)和束缚态淀粉合成酶(GBSS)活性在籽粒发育过程中一直最高,有利于直链淀粉、支链淀粉的积累和支链淀粉/直链淀粉比的提高,提高峰值粘度、低谷粘度、最终粘度和衰减值;花后土壤含水量过高(80%～90%)或过低(40%～50%)均导致籽粒SSS和GBSS活性降低,从而使直链淀粉、支链淀粉的积累量降低,减小支链淀粉/直链淀粉比,使峰值粘度、低谷粘度、最终粘度降低.(2)在同一土壤含水量下,增施氮肥不利于灌浆前期SSS和GBSS活性和直链淀粉、支链淀粉积累量的提高,并且随着土壤含水量增加增施氮肥该趋势加重;适量增施氮肥能提高支链淀粉/直链淀粉比和峰值粘度、低谷粘度、最终粘度,过多或过少施氮则降低支/直比和峰值粘度、低谷粘度、最终粘度.研究认为,在本试验条件下,适量增施氮肥(纯氮225 kg/hm2)或适宜的花后土壤含水量(60%～70%)可促进强筋小麦籽粒淀粉的合成,有效改善其淀粉品质.     

应用CRISPR/Cas9技术创制低直链淀粉含量水稻种质

Wx基因直接控制水稻胚乳直链淀粉的合成,进而影响胶稠度等多个品质指标,是控制稻米食味的主效基因.适当降低稻米的直链淀粉含量可改善稻米品质.吉丰B是籼型优质杂交稻保持系,直链淀粉含量偏高,影响组配后代的食味品质.本研究以吉丰B为试验材料,对Wx基因编码区上游序列进行编辑,以期获得Wx基因表达下调,直链淀粉含量降低的水稻种...     

水稻SSIIa基因型对成糊温度校准的影响及糊化温度与直链淀粉含量的关系分析

稻米淀粉黏滞性谱(RVA谱)常被用来鉴定水稻种质资源和所选优良株系的食味品质,RVA成糊温度(PT)却不能准确反映糊化温度(GT),校准成糊温度PT m能够比较准确地反映GT,但PT的校准范围、PT m衡量稻米GT的准确程度以及SSIIa基因型对其的影响等并未见报道。本研究利用遗传背景差异小、双亲糊化温度差异大的B1F8株系中的58个单株为材料,通过检测其SSIIa基因型,将材料分为3类,分别在3种SSIIa基因型的遗传背景下,比较分析PT、PT m、GT,结果表明,SSIIa基因型不同,PT的校准范围明显不同;PT衡量稻米GT的准确率仅为39.66%,而PT m准确率达到89.66%。进一步利用PT和PT m分别表示GT,分析稻米AC与GT关系,得出与一些学者不同的结论:稻米AC与GT呈极显著负相关性。     

玉米种子发育过程中直链淀粉积累规律的分析

淀粉是玉米种子的主要组成成分,它包括直链淀粉和支链淀粉。支链淀粉的合成需要淀粉合成酶、分支酶和脱支酶的共同作用,而直链淀粉的合成则是在颗粒结合型淀粉合成酶的作用下进行的。颗粒结合型淀粉合成酶基因的突变造成玉米种子的腊质（糯性）表型。与支链淀粉合成的分子机制的研究相比,目前对玉米种子中直链淀粉合成的分子机制了解相对较少。以野生型黄早4玉米自交系和突变体糯玉米为实验材料,研究了种子不同发育时期直链淀粉的积累规律。通过碘染色的方法,观察了玉米种子发育过程中淀粉积累的形态变化。定量分析表明,从授粉后10d至25d,黄早4种子中直链淀粉的含量逐渐增加,同时颗粒结合型淀粉合成酶（GBSS）的活性逐渐提高;而在糯玉米中,直链淀粉和GBSS活性均未检测到。进而,通过RT-PCR方法,从黄早4种子中分离出编码GBSSI的cDNA片段。在授粉后10d至25d的玉米胚乳中均可检测到GBSSI的表达,而在胚中直到授粉后25d才检测到该基因表达的微弱信号。在糯玉米种子中没有检测到该基因的表达。研究结果表明,在玉米种子发育过程中,GBSSI基因的表达通过控制GBSS的合成,最终控制直链淀粉的合成。研究工作为理解玉米种子中直链淀粉合成的分子机制提供了重要信息。     

水、旱条件下稻米蒸煮和营养品质性状与土壤水分环境互作分析及其QTL定位

以旱稻品种IRAT109与水稻品种越富杂交构建的DH群体的116个株系及其亲本为材料,在水、旱2种栽培条件下种植,研究了稻米蒸煮和营养品质性状的变化规律,在水、旱2个土壤水分环境下对直链淀粉含量(AC)、胶稠度(GC)、碱消值(GT)和蛋白质含量(PC)4个蒸煮和营养品质性状进行QTL定位及QTLs与环境互作分析。结果表明,以上4个品质性状在水、旱2种不同栽培条件下差异较大,说明这些性状受水分条件影响较大,旱栽条件下稻米蒸煮和营养各品质性状均有不同程度的升高,其中蛋白质含量平均提高37.9%。QTL分析结果表明,4个稻米品质性状在2个环境中的表现型值都为连续分布,均存在超亲遗传类型,共检测到7个加性效应QTL与稻米蒸煮和营养品质性状4项指标有关,分别位于第1、2、3、6、8、11染色体上,单个QTLs对性状的贡献率在1.91%～19.77%之间。位于第3染色体上控制碱消值的QGt3,第6染色体上控制直链淀粉含量的QAc6,在2个不同土壤水分条件下均与环境存在显著互作,对环境互作的贡献率分别为8.99%和47.86%。控制直链淀粉含量的2对上位性QTLs与土壤水分环境显著互作,贡献率较大,分别为32.54%和11.82%。并筛选到5个主效QTL(QGt6b、QGt8、QGt11、QGc1和QPc2)在抗旱育种中可用于蒸煮和营养各品质性状MAS改良。     

Field Performance of Transgenic indica Hybrid Rice with Improved Cooking and Eating Quality by Down-regulation of Wx Gene Expression

High amylose content (AC) in rice endosperm is correlated with poor grain quality, particularly in indica hybrid rice. We have generated several homozygous transgenic parent lines of indica hybrid rice carrying an antisense Waxy (Wx) gene and demonstrated that the AC in seeds of these lines decreased dramatically. Two transgenic maintainer lines (L25B and L18B), derived from one of the key maintainer parents of an indica hybrid rice in China, Long-te-fu B (LTF-B), were selected and the antisense Wx gene was subsequently introgressed into the male-sterile counterpart, LTF-A, with the aim to generate improved indica hybrids. The indica hybrids derived from the selected transgenic male-sterile lines and restorer lines were tested for quality and agronomic performance under normal field conditions. Our results demonstrated that the reduction of AC in the homozygous transgenic maintainer lines stably passed down in five successive generations and the improved quality was also found in their relevant transgenic hybrids produced. The other two key characters of rice cooking and eating quality, the gel consistence (GC) and gelatinization temperature (GT), were also improved in the grains of both the transgenic maintainer lines and their relevant hybrids. In addition, no change was observed for most of the agronomic characters of the transgenic maintainer lines and the relevant transgenic hybrids. Although the grain weight of the transgenic line was reduced, the grain yield of the homozygous transgenic parent lines and the transgenic hybrids was similar when compared with that of the wild-type controls. These results suggest that transgenic approaches are an effective way to obtain rice lines with both improved qualities and high yield, especially for indica hybrid rice.     

优质稻米‘青香软粳’低直链淀粉含量形成分子机制的初步研究

Waxy（Wx）基因在调控直链淀粉含量形成过程中起着重要的作用,是稻米蒸煮食味品质的一个关键决定因素。在本项研究中,我们以直链淀粉含量为8.7%和10.2%的优质稻品种‘青香软粳’和‘南粳46’为主要研究对象,初步分析并探讨了这两个优质稻品种稻米低直链淀粉形成的分子调控机理。结果显示,虽然都为Wx-II型粳稻品种,‘南粳46’的Wx基因在启动子1773位置存在AA/CT的变异,‘南粳46’和‘青香软粳’的Wx基因在启动子＋693位置具有G/A位点多态性。转录水平分析表明,在‘南粳46’和‘青香软粳’稻米发育过程中,Wx基因的表达模式有较大的差异,可能与其启动子序列的多态性相关,AA/CT变异处于Wx基因的转录调控区。Wx基因启动子后＋693位点的多态性与已报道Wxmq多态性位点之一相同,引起Arg158/His158的变异;生物信息学软件分析表明,该位点处于底物进入Wx蛋白催化中心的袋口上,Arg158/His158位点的变异可能影响到底物进入Wx蛋白催化活性中心的速率,从而影响稻米直链淀粉的合成过程和含量。文章丰富了Wx基因多态性的研究,为进一步验证其核苷酸变异与稻米直链淀粉含量的相关性奠定了基础。     

Genetic basis of 17 traits and viscosity parameters characterizing the eating and cooking quality of rice grain

A recombinant inbred line population derived from a cross between Zhenshan 97 and Delong 208 was used to analyze the genetic basis of the cooking and eating quality of rice as reflected by 17 traits (or parameters). These traits include amylose content (AC), gel consistency (GC), alkali spreading value (ASV), cooked rice elongation (CRE), and 13 parameters from the viscosity profile. All the traits, except peak paste viscosity (PKV), time needed from gelatinization to peak (BAtime), and CRE, can be divided into two classes according to their interrelationship. The first class consists of AC, GC, and most of the paste viscosity parameters that form a major determinant of eating quality. The second class includes ASV, pasting temperature (Atemp) and pasting time (Atime), which characterize cooking process. We identified 26 QTL (quantitative trait locus or loci) in 2 years; nine QTL clusters emerged. The two major clusters, which correspond to the Wx and Alk loci, control the traits in the first and second classes, respectively. Some QTL are co-located for the traits belonging to the same class and also for the traits to a different class. The Wx locus also affects on ASV while the Alk locus also makes minor contributions to GC and some paste viscosity parameters. The QTL clusters on other chromosomes are similar to the Wx locus or Alk locus, although the variations they explained are relatively minor. QTL for CRE and PKV are dispersed and independent of the Wx locus. Low paste viscosity corresponds to low AC and soft gel, which represents good eating quality for most Chinese consumers; high ASV and low Atemp, together with reduced time to gelatinization and PKV, indicate preferred cooking quality. The genetic basis of Atemp, Atime, BAtime, peak temperature, peak time, paste viscosity at 95 degrees C, and final paste viscosity is newly examined to reveal a complete and dynamic viscosity profile.     

Strategy and utilization of a herbicide resistance gene in two-line hybrid rice

Hybrid rice plays an important role in China's aim to improve rice production as it accounts for some 50% of rice planting area but produces about 60% of the total rice grain. However, the existing three-line system used in hybrid rice production has its limitations. The two-line system, which makes use of photoperiod-sensitive genic male-sterile (PGMS) and thermo-sensitive genic male-sterile (TGMS) lines to generate the male-sterile parental line, was developed to overcome some of these limitations. The sterility of the male-sterile line of two-line hybrid rice, however, fluctuates when the temperature-sensitive phase of fertility encounters abnormal low temperatures during hybrid seed production, which induces selfing and decreases the purity of hybrid. We describe here the strategy of utilizing a herbicide resistance gene in two-line hybrid rice to eliminate this fluctuation in the sterility of the P/TGMS lines during hybrid seed production and reports the development of the herbicide resistance restorer line Bar68-1 and its herbicide-resistant early season hybrid rice Xiang125s/Bar68-1. When the restorer line and its derived hybrid are herbicide resistant, the selfed seeds can be removed easily from the hybrid by herbicide spraying. A herbicide resistance gene bar was transferred into a restorer line by particle bombardment. The resulting transgenic restorer line Bar68-1 and its hybrid Xiang125 s/Bar68-1 inherited stable herbicide resistance. The purity of Xiang125s/Bar68-1 was increased by spraying the seed bed with herbicide, which resulted in a significant increase in yield, grain quality, and disease resistance in comparison to the controls in a regional trial.     

A new AOX homologous gene OsIM1 from rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) with an alternative splicing mechanism under salt stress

"Zhenshan 97" is the female parent of a number of widely used hybrids for rice production in China. However, this line is of poor quality because of a high amylose content (AC), a hard gel consistency (GC) and a low gelatinization temperature (GT), together with a chalky endosperm. It had been determined that the three traits for cooking and eating quality, AC, GC and GT, are controlled by the Waxy locus and/or the tightly linked genomic region. In this study we improved the eating and cooking quality of Zhenshan 97 by introgressing the Waxy gene region from Minghui 63 (wx-MH), a restorer line, that has medium AC, soft GC and high GT. The wx-MH fragment was transferred to Zhenshan 97B by three backcrosses and one selfing, then from Zhenshan 97B to Zhenshan 97A by a cross and a backcross. Molecular marker-assisted selection was applied in the series to select for individuals carrying wx-MH, to identify recombination between the Waxy and flanking markers, and also to recover the genetic background of the recurrent parent. According to the marker genotypes, the improved versions of Zhenshan 97B and Zhenshan 97A, or Zhenshan 97B(wx-MH) and Zhenshan 97A(wx-MH), were the same as the originals except for the Waxy region of less than 6.1 cM in length. The selected lines and their hybrids with Minghui 63, or Shanyou 63(wx-MH), showed a reduced AC and an increased GC and GT, coupled with a reduced grain opacity. Field examinations of agronomic performance revealed that Zhenshan 97B(wx-MH) and Shanyou 63(wx-MH) were essentially the same as the originals except for a significant decrease in grain weight. The simultaneous improvement of AC, GA, GT and opacity, indicated that the Waxy region had major effects on the four quality traits. The improved versions of Zhenshan 97 A and B should be immediately useful in hybrid rice production.     

籼爪交水稻F_2群体的蒸煮食味品质研究

分析了籼型高直链淀粉恢复系CG133R与糯性爪哇稻22号杂交衍生的F2群体的蒸煮品质变异及其与淀粉粘滞性特征间的相关性,以及F2群体颗粒结合淀粉合成酶(Wxa基因)和可溶性淀粉合成酶(SSⅡ-3基因)主效基因的遗传。结果表明:蒸煮品质指标和RVA谱特征值在F2群体中广泛分离,其中变异最大的是消减值,其次为胶稠度、直链淀粉含量。高直链淀粉材料各理化指标与RVA谱特征值的相关性不显著;RVA谱特征值在中、低直链淀粉含量和糯稻群体中与各理化指标存在显著或极显著相关;在中、低直链淀粉材料中,RVA谱特征值与糊化温度(GT)也存在显著或极显著相关。用Wxa基因和SSⅡ-3基因的分子标记检测到这两个基因在F2群体中存在偏分离,分别指向两个亲本类型。除高直链淀粉材料外,可以通过RVA谱特征值来辅助筛选优质水稻品种。     

Novel Wx alleles generated by base editing for improvement of rice grain quality

Amylose content (AC), which is regulated by the Waxy (Wx) gene, is a major indicator of eating and cooking quality (ECQ) in rice (Oryza sativa). Thus far, only a limited number of mutations in the N-terminal domain of Wx were found to have a major impact on the AC of rice grains and no mutations with such effects were reported for other regions of the Wx protein. Here, nucleotide substitutions in the middle region of Wx were generated by adenine and cytosine base editors. The nucleotide substitutions led to changes in 15 amino acid residues of Wx, and a series of novel Wx alleles with ACs of 0.3%–29.43% (wild type with AC of 19.87%) were obtained. Importantly, the waxy^abe2 allele showed a “soft rice” AC, improved ECQ, favorable appearance, and no undesirable agronomic traits. The transgenes were removed from the waxy^abe2 progeny, generating a promising breeding material for improving rice grain quality.     

中籼杂交水稻亲本多态性的AFLP分析

对15个籼型杂交水稻亲本进行AFLP分析,结果表明：亲本间遗传距离小,在0.0589-0.3305之间,平均为0.2033。15个亲本按类平均法可聚为两类,Ⅰ类为不育系,Ⅱ类为恢复系。其中Ⅱ类又分为两个亚类,Ⅱ－1不含明恢63血缘、Ⅱ－2全部含明恢63血缘。Ⅰ/Ⅱ－1与Ⅰ/Ⅱ－2间的遗传距离无明显差异,揭示恢复系的遗传基础较一致,这可能是当前的品种不能超过籼优63的重要原因之一。要提高水稻的杂种优势,需丰富亲本的遗传基础,扩大其遗传差异。     

水稻红莲型细胞质雄性不育系小孢子发育过程中的花药蛋白质分析(英文)

水稻不育系的不育机理至今尚未搞清。本工作对红莲型不育系小孢子发育过程中花药蛋白质的变化进行了分析,为从分子水平阐明其不育机理奠定基础。分别在不育系（A）、保持系（B）、杂种一代（F1）和恢复系（R）的花粉母细胞单核、二核及三核四分体时期取花药,匀浆、反复离心制备上清液、定量后,做不连续Bis－Acr凝胶电泳、染色,观察其蛋白电泳条带的变化。结果见Fig．1＆2和Tab．1：大多数主要的、表达量大的蛋白质在二核期前后（花粉败育时期）,没有明显变化,这些蛋白与败育无关。条带X、XI随着小孢子发育过程出现变化,但它们在A、B、F1和R中的表达行为变化一致,也与败育无关。条带Ⅰ、Ⅱ在不育系中不表达,条带Ⅲ为不育系所特有,条带Ⅳ在单核期以后出现,条带Ⅶ在二核期受到抑制,它们均可能与育性有关。A、F1中均有条带Ⅴ表达,说明该编码基因是由不育系遗传到F1中的,它与育性无关;而R、F1系中均有条带Ⅵ、Ⅷ表达,该编码基因可能是由恢复系遗传到F1中的,可能涉及F1的育性恢复。