旨在找到一种简便的SNP检测方法用于水稻种性鉴定及遗传多样性分析,本研究以245份长江中下游主推杂交水稻品种及亲本为研究材料,从1319个SNP位点中筛选出124对多态性高的位点,建立SNP的高效检测体系.选用其中的1个标记sf0141821553和SSR的标记RM7120来检测水稻的纯度,分别利用Caps-AccI标记和SNP-sf0601764762扩增亲本及杂交后代Wx的基因型,两者结果完全一致.用124对位点对245份杂交水稻进行遗传多样性分析,聚类分析显示,245个品种间的遗传相似系数在0.64～0.97之间.以遗传相似系数0.64为阈值,可以将245个水稻品种分为2个类群,这2个类群分别在遗传相似系数为0.712和0.667处又可以分为2个亚群.结果 表明材料间存在明显的群体结构,能精确区分待测品种的基因型以及品种间的遗传差异.利用聚丙烯酰胺凝胶电泳来实现SNP标记的检测,方法简便且不需要大型仪器设备和昂贵的试剂,便于实验室开展水稻品种的鉴定工作.

This study aims to solve the problem of distinguishing wheat varieties.160 main cultivated wheat varieties were taken as experimental materials from Huang-Huai-Hai region and the middle-to-lower reaches of the Yangtze River,21 pairs of stable and high polymorphism primers on each chromosome were screened from 21 linkage groups distributed in 105 pairs of SSR primers as SSR fluorescence based on capillary electrophoresis analysis.The authors constructed the wheat variety fingerprints and digitally encoded molecular ID of 160 wheat varieties combining with the varieties' basic commodity information.The results realize the unity of the uniqueness,universality,identifiability and traceability of wheat varieties' information,which provide convenience for the quality tracing and management of wheat seeds and scientific evidence for the protection of intellectual property right.

Colour and moisture content are important indices in quality monitoring of dehydrating carrot slices during dehydration process. This study investigated the potential of using multispectral imaging for real-time and non-destructive determination of colour change and moisture distribution during the hot air dehydration of carrot slices. Multispectral reflectance images, ranging from 405 to 970 nm, were acquired and then calibrated based on three chemometrics models of partial least squares (PLS), least squares-support vector machines (LS-SVM), and back propagation neural network (BPNN), respectively. Compared with PLS and LS-SVM, BPNN considerably improved the prediction performance with coefficient of determination in prediction (RP(2))=0.991, root-mean-square error of prediction (RMSEP)=1.482% and residual predictive deviation (RPD)=11.378 for moisture content. It was concluded that multispectral imaging has an excellent potential for rapid, non-destructive and simultaneous determination of colour change and moisture distribution of carrot slices during dehydration.

BACKGROUND:Rancidity is an important attribute for quality assessment of butter cookies, while traditional methods for rancidity measurement are usually laborious, destructive and prone to operational error. In the present paper, the potential of applying multi-spectral imaging (MSI) technology with 19 wavelengths in the range of 405-970 nm to evaluate the rancidity in butter cookies was investigated.RESULTS:Moisture content, acid value and peroxide value were determined by traditional methods and then related with the spectral information by partial least squares regression (PLSR) and back-propagation artificial neural network (BP-ANN). The optimal models for predicting moisture content, acid value and peroxide value were obtained by PLSR. The correlation coefficient (r) obtained by PLSR models revealed that MSI had a perfect ability to predict moisture content (r = 0.909), acid value (r = 0.944) and peroxide value (r = 0.971).CONCLUSION:The study demonstrated that the rancidity level of butter cookies can be continuously monitored and evaluated in real-time by the multi-spectral imaging, which is of great significance for developing online food safety monitoring solutions.

In order to prevent the emergence of “forged and fake commodity” and “fleeing goods” in wheat seed industry, strengthen the industry security, and improve the level of intelligent supervision and management, we proposed a new coding scheme related to the identity of wheat seed to generate a new traceability code for tracking based on the existing traceability technology and smart packaging. Furthermore, we designed and implemented an analysis model for anti-fake and forecasting, which can intelligently show managers and customers the areal distribution and influence degree of the fake wheat seed. By using the traceability code and analysis model, managers and customers can clearly determine the authenticity of the wheat seed. Our results showed that the new traceability code and analysis model have significant efficiency in traceability, anti-fake and forecasting of wheat seed, indicating their great practical value and application prospect for the wheat seed industry.

Total polyphenols is a primary quality indicator in tea which is consumed worldwide. The feasibility of using near infrared reflectance (NIR) spectroscopy (800–2500 nm) and multispectral imaging (MSI) system (405–970 nm) for prediction of total polyphenols contents (TPC) of Iron Buddha tea was investigated in this study. The results revealed that the predictive model by MSI using partial least squares (PLS) analysis for tea leaves was considered to be the best in non-destructive and rapid determination of TPC. Besides, the ability of MSI to classify tea leaves based on storage period (year of 2004, 2007, 2011, 2012 and 2013) was tested and the classification accuracies of 95.0% and 97.5% were achieved using LS-SVM and BPNN models, respectively. These overall results suggested that MSI together with suitable analysis model is a promising technology for rapid and non-destructive determination of TPC and classification of storage periods in tea leaves.

为准确追溯、快速鉴定种子质量,保证种子的质量安全,从农作物品种身份证的组成、农作物种子电子代码的实现、农作物种子电子代码的应用以及存在的问题等方面总结了农作物种子电子代码的应用情况.

为解决种子生产、经营和知识产权保护中存在的突出问题,本文提出了构建农作物品种身份证及建立品种身份证制度.在借鉴人类身份证建立模式基础上,将品种的商品信息、指纹信息及特异基因信息相结合,构建农作物的品种身份证.农作物品种身份证由3部分构成:第1部分称为商品码,反映作物及品种类别、品种选育(或审定)的区域和时间等,预设14位阿拉伯数字(或字母);第2部分称为指纹码,反映品种的DNA指纹信息,依据作物类型不同预设20～40位阿拉伯数字(或字母);第3部分称为补充码(特异基因识别码),反映品种的特异基因信息,设计为字母“T”(转基因育种)、“S”(分子育种)和“M”(诱变育种)等,加上被导入、渗入或诱变的基因名称;身份证总位数约50位.采用本方法编码的农作物品种身份证,具有准确性高、通用性强及可追溯性等特点,对我国农作物品种的科学识别和管理具有十分积极的意义.

利用分布于油菜连锁群上的33对SSR核心引物,对12个育种品系和市场上的杂交油菜品种进行鉴定,探讨取样方案、检测方法和位点判读,将样品间SSR位点类型分为相同位点、疑似相同位点和差异位点3类.分析位点类型发现,12个油菜品种/系间的差异位点数为4 ～ 22,其中同父异母杂交种(3个)间差异位点数为4～16,同母异父杂交种(4个)间差异位点数为7 ～18,市场抽样的品种间差异位点数为12 ～ 20.同一品种不同样本间的差异位点数为0～2,据此,将区分品种间与品种内的差异位点阈值设定为2,即在利用33对引物对油菜杂交种与对照样品进行SSR分析时,当两样品间差异位点数＞2时,判定为不同品种;当差异位点数≤2时,判定为近似或极近似品种.盲样鉴定实验及其与田间鉴定方法的对比实验,均证明了本研究确定的SSR鉴定方法可行.

Multispectral imaging in the visible and near-infrared (405–970 nm) regions was tested for nondestructive discrimination of insect-infested, moldy, heterochromatic, and rancidity in sunflower seeds. An excellent classification (accuracy >97 %) for intact sunflower seeds could be achieved using Fisher’s linear discriminant function based on 10 feature wavelengths that were selected from the original 19 wavelengths by Wilks’ lambda stepwise method. Intact sunflower seeds with different degree of rancidity could be precisely clustered by multispectral imaging technology combined with principal component analysis-cluster analysis (PCA-CA). Our results demonstrate the capability of multispectral imaging technology as a tool for rapid and nondestructive analysis of seed quality attributes, which enables many applications in the agriculture and food industry.

Crop-to-crop transgene flow may affect the seed purity of non-transgenic rice varieties, resulting in unwanted biosafety consequences. The feasibility of a rapid and nondestructive determination of transgenic rice seeds from its non-transgenic counterparts was examined by using multispectral imaging system combined with chemometric data analysis. Principal component analysis (PCA), partial least squares discriminant analysis (PLSDA), least squares-support vector machines (LS-SVM), and PCA-back propagation neural network (PCA-BPNN) methods were applied to classify rice seeds according to their genetic origins. The results demonstrated that clear differences between non-transgenic and transgenic rice seeds could be easily visualized with the nondestructive determination method developed through this study and an excellent classification (up to 100% with LS-SVM model) can be achieved. It is concluded that multispectral imaging together with chemometric data analysis is a promising technique to identify transgenic rice seeds with high efficiency, providing bright prospects for future applications.

本研究以抗虫水稻恢复系9311（Bt）及其杂交种F1（Bt）为研究材料,以非转基因的9311为阴性对照,利用ELISA方法研究9311（Bt）及F1（Bt）各生育时期可溶性总蛋白和Bt蛋白的时空变化规律,为转Bt基因抗虫水稻的安全监管提供科学依据。结果表明:外源基因的导入没有引起水稻组织中可溶性总蛋白含量的明显变化;9311（Bt）的Bt蛋白表达量在整个生长周期的各个部位均高于相应的F1（Bt）植株;同一植株不同组织器官中Bt蛋白表达量为:叶片＞胚乳＞颖壳及茎秆＞根;同一植株不同发育期叶片Bt蛋白的测定结果整体表现为:营养生长阶段＞生殖生长阶段＞成熟衰老阶段。研究结果为转Bt基因抗虫水稻适宜检测时期的选择提供了一定的参考。

Multispectral imaging with 19 wavelengths in the range of 405-970 nm has been evaluated for nondestructive determination of firmness, total soluble solids (TSS) content and ripeness stage in strawberry fruit. Several analysis approaches, including partial least squares (PLS), support vector machine (SVM) and back propagation neural network (BPNN), were applied to develop theoretical models for predicting the firmness and TSS of intact strawberry fruit. Compared with PLS and SVM, BPNN considerably improved the performance of multispectral imaging for predicting firmness and total soluble solids content with the correlation coefficient (r) of 0.94 and 0.83, SEP of 0.375 and 0.573, and bias of 0.035 and 0.056, respectively. Subsequently, the ability of multispectral imaging technology to classify fruit based on ripeness stage was tested using SVM and principal component analysis-back propagation neural network (PCA-BPNN) models. The higher classification accuracy of 100% was achieved using SVM model. Moreover, the results of all these models demonstrated that the VIS parts of the spectra were the main contributor to the determination of firmness, TSS content estimation and classification of ripeness stage in strawberry fruit. These results suggest that multispectral imaging, together with suitable analysis model, is a promising technology for rapid estimation of quality attributes and classification of ripeness stage in strawberry fruit.

利用均匀分布于棉花26条染色体上的39对SSR核心引物对16个不同来源的棉花品种样品进行区别与鉴定实验,探讨了棉花品种鉴定的取样方案和位点异同判读方法.通过比较不同取样量样品包含的遗传信息,确定了基本的取样量,并将样品间位点异同类型分为相同位点、疑似相同位点和差异位点.据此对16个品种进行位点异同性分析,结果表明同一品种的不同样品间差异位点数均在0～2间,而不同品种间的差异位点数介于5～25.结合品种间差异位点的阈值分析,研究认为当两样品间差异位点数＞2时,即判定为不同品种;当差异位点数≤2时,即判定为近似或极近似品种.

On the base of the previous research, we selected a set of 12 rice SSR markers(one marker for each chromosome) for increasing convenience of the rice seed quality tracking and management. The fluorescent marker analysis was performed on 127 released and popularized authorized(or main plant) rice varieties in Anhui Province. Then, the rice variety ID was constructed by the SSR fingerprint with corresponded commodity information. The rice variety ID included three components: part 1 was recognized as commodity code, which reflected crop species, cultivar, breeding(or approval) region and year; part 2 was recognized as fingerprint code, which contained the variety DNA fingerprint information; part 3 was recognized as complement code(specific gene identification code), reflecting specific genetic information of cultivars. We also described the rice variety ID in the form of bar code or QR code, which might be favorable for rice seed identification and traceability management.

Germplasm accurate identification is usually affected by the similar phenotypic characteristics performed in the field,it is necessary to establish ID to distinguish varieties.463 SSR primers covering 19 linkage groups of rapeseed chromosome were screened,33 primers showed polymorphic bands with 191 alleles.The number of alleles by each primer was 3 to 9 with an average of 5.787 9 per primer,the effective number of alleles(Ne) is 1.906 8 to 7.479 0 with an average of 4.730 7,the diversity(Nei's genetic diversity index,He) was 0.773 8 to 2.045 1 with an average of 1.551 5,the discrimination power(DP) was 0.389 2 to 0.862 9 with an average of 0.742 8,polymorphism index content(PIC) was 0.475 6 to 0.866 3 with an average of 0.764 0.The principal coordinate(PCO) analysis indicated that all material could be clearly distinguished by 33 pairs of core primers,which divided into 3 subfamiles based on genetic similarity.After gradually screening,7 pairs core primer(BrGMS075,Na14E08,BrAS084,CB10545,Na14G10,CN57 and Ra2E04) were selected,and their bands amplified by each marker were coded,and then the codes were combined as a molecular ID for Brassica napus.

In this study,we performed SSR(simple sequence repeats) analysis of 133 hybrid parental rice varieties with 48 SSR core primer pairs recommend by GB/T20396-2006.Based on polymorphism among the materials,we built molecular ID and fingerprint for each variety.The ID was constructed by 48 digit numbers and letters,which represent the detail polymorphism on certain variety.Combined the SSR polymorphism and anatomical information,a searchable SSR fingerprinting database was built.This database could be used to identify the basic information,molecular ID and similarity of varieties.Furthermore,it also could be applied in seed purity examining,parental similarity testing to support the rice hybrid breeding.This database not only sets a sharing platform for examination institutes or breeders,but also technically supports the breeding,testing,managing and judging on rice varieties.

Two rice varieties with similar apparent amylose content, Nipponbare (Oryza sativa subsp. japonica) and 93-11 (O. sativa supbsp. indica), were used as parents to establish a recombinant inbred line population consisting of 190 lines by single seed descent method. The genetic linkage map contained 202 simple sequence repeat (SSR), cleaved-amplified polymorphism sequence (CAPS), and sequence tagged site (STS) markers. Quantitative trait loci (QTLs) were identified for 8 rice starch RVA profile properties including peak paste viscosity (PKV), hot paste viscosity (HPV), cool paste viscosity (CPV), breakdown viscosity (BDV), setback viscosity (SBV), consistence viscosity (CSV), peak time (PeT), and pasting temperature (PaT) in 3 ecological sites using composite interval mapping method. A total of 57 QTLs were identified, with 1 to 14 for each trait. Thirteen stable QTLs were detected at repeated environments, among which qCPV-3, qCPV-10b, qSBV-10b, qCSV-3b, and qCSV-10b were detected in all environments, and they explained 26.9%, 29.5%, 29.7%, 25.2%, and 28.3% of phenotypic variations, respectively. Sixteen QTLs had pleiotropy with a single QTL controlling 2–6 RVA profile properties. The interval of RM25032-RM1375 on chromosome 10 harbored gene loci controlling PKV, HPV, CPV, SBV, PaT, and CSV.

To improve the Agrobacterium-mediated transformation frequency of Japonica rice Wanjing 97,an orthogonal experiment L9(33) was designed to optimize the factors regulating rice genetic transformation,and the impacts of innoculum OD600,coculture temperature and coculture duration on transformation using gfp gene to monitor the expression of target gene were analyzed.Results showed that the optimal conditions for Agrobacterium-mediated transformation of Wanjing 97 were determined to be innoculum OD660 of 0.1,cocultured at 21℃-23℃ dark for 72 h.Under this condition,the rate of gfp-positive calli was up to 70.9%.The results suggested that gfp could be used as an effective reporter gene to improve rice genetic transformation.