含不同Anti-Waxy基因拷贝数的稻米直链淀粉含量分析

将含单拷贝Anti-Waxy基因的纯合植株分别作为父本或母本进行正反交,获得两组含双拷贝Anti-Waxy基因的杂交后代,分析了未转基因对照、转基因亲本及两组杂交后代糙米直链淀粉含量以揭示不同Anti-Waxy基因拷贝数对降低稻米直链淀粉含量程度的影响.结果显示,2个单拷贝转基因水稻糙米直链淀粉平均含量分别为10.72%和11.13%,比对照分别降低17.98%和14.84%;两组正交和反交杂交后代糙米直链淀粉平均含量分别为8.96%和8.23%,其平均值为8.60%,比2个转基因杂交亲本糙米直链淀粉含量分别降低19.78%和22.73%,比对照降低了34.20%.结果表明,增加转基因水稻基因组中Anti-Waxy基因拷贝数在一定程度上能够进一步降低稻米直链淀粉含量,通过将独立转化获得同品种转基因植株之间杂交可以成为获得高表达转基因植物的途径.     

导入反义蜡质基因改良水稻稻米的食味品质和营养品质

经PCR、Southern blot和稻米GUS检测,蜡质基因反义片段与gus构成的融合基因已整合到8株水稻基因组中,并能正确表达。通过稻米GUS染色追踪分析获得转反义蜡质基因纯合后代。将转基因植株的稻米送农业部稻米及制品质量监督检验测试中心进行糙米蛋白质含量和直链淀粉含量测定。结果显示：（1）在转反义蜡质基因纯合的超2-10粳稻后代中,大多数单株糙米在直链淀粉含量降低的同时,蛋白质含量有不同程度提高,糙米蛋白质含量和直链淀粉含量呈显著负相关关系;（2）对照糙米直链淀粉平均含量和糙米蛋白质平均含量分别为13.4%和9.5%。在转基因植株中,稻米直链淀粉含量最低的为11.4%,而蛋白质含量也相对最高,为13.5%。本试验结果表明在水稻中导入反义蜡质基因不仅能够降低水稻稻米的直链淀粉含量,还有可能提高水稻稻米的蛋白质含量。     

转反义蜡质基因‘湘晴’及其杂交稻米的直链淀粉含量研究

利用根癌农杆菌介导将反义蜡质基因(anti-Waxy)导入三系恢复系湘晴水稻获得转基因植株.从T1代外源基因呈单拷贝整合的转基因湘晴水稻中选取3个稻米直链淀粉含量降低较明显的单株继续种植得到纯合后代.以纯合转基因植株及湘晴水稻(对照)为恢复系分别与寒丰不育系杂交,获得4组杂交稻后代(F2).3个T3代纯合转基因湘晴水稻糙米(T4)直链淀粉含量分别为14.42%、13.96%和14.72%,对照为16.04%;3组由转基因湘晴水稻为父本杂交制种后代(F2)糙米直链淀粉含量分别为14.53%、13.77%和14.64%,对照杂交稻(F2)为16.22%.研究表明,导入湘晴水稻中的反义蜡质基因不仅能够降低湘晴稻米直链淀粉含量,还能够进一步抑制杂交后代的稻米直链淀粉合成.     

籼稻品质分析的近红外光谱模型建立及其应用研究

为了满足籼稻品质快速分析的需求,本研究利用籼稻精米粉近红外光谱建立了直链淀粉含量、蛋白质含量、碱消值、垩白度的回归预测模型.结果表明,本研究提供的预测模型具有良好的测定效果,用偏最小二乘法(PLS)获得的籼稻精米粉直链淀粉含量、蛋白质含量、碱消值、垩白度的回归模型和交叉验证显示最优校正决定系数(R~2)和交叉检验均方误差(RMSECV)分别为0.9561、1.55,0.9510、0.258,0.9076、0.283,0.9014、4.14.说明所建的近红外光谱预测模型具有实用价值.     

绿豆主要营养品质近红外预测模型的构建与验证

运用近红外法预测了中国绿豆主要营养品质蛋白质、淀粉和直链淀粉含量并比较绿豆粉和籽粒两种不同样品类型的差异。102份来自绿豆核心样品为试验材料,采用近红外分析系统扫描了绿豆籽粒和粉样品。光谱数据经预处理后,构建了最小二乘回归预测和交互验证模型并获得了最优校正统计参数。绿豆粉样品最大R2值和最小SECV值蛋白质含量为0.95和0.329,淀粉含量0.90和0.576,直链淀粉含量0.89和0.307;RPD值3.08至4.61。籽粒样品最大R2值和最小SECV值蛋白质含量为0.90和0.404,淀粉含量0.88和0.643,直链淀粉含量0.85和0.426;RPD值2.51至3.23。模型的稳健性采用外部验证法进行了评价。豆粉样品的平均差异1.0%～1.8%比籽粒样品略低。结果表明绿豆粉的三种组分的近红外预测方法具有快速和简单的特点,可应用于绿豆品质的测定。籽粒样品还具有无损种子结构,保持种子活力的特点,可在育种、种质资源创新等方面应用,但其准确度还有待进一步的改善。     

水稻籽粒直链淀粉含量的生态模型研究

在中国、日本和泰国不同生态环境下进行了多品种籼稻和粳稻的种植试验,通过分析水稻籽粒直链淀粉含量与纬度、海拔、温度、太阳辐射等气候生态因子的相互关系,确立了影响水稻籽粒直链淀粉积累的主要气候因子函数,并使用权重系数进一步修订各个气候生态因子对籽粒直链淀粉的作用,构建出基于生态效应(综合气候因子函数)的水稻籽粒直链淀粉含量预测模型.利用不同年份、不同生态点和不同品种类型的试验资料对所建模型进行了检验,籼稻和粳稻籽粒直链淀粉含量的预测误差(RMSE)平均分别为0.4%和0.5%;籼稻和粳稻种植区的预测误差(RMSE)平均为0.39%和0.50%,表明模型具有较好的预测性和实用性.     

臭氧胁迫对稻穗不同部位糙米直链淀粉含量和RVA谱特征值的影响

近地层臭氧浓度升高使水稻产量下降,但臭氧胁迫对稻米品质的影响及其强弱势粒差异尚不清楚.本研究以8个不同类型水稻品种为供试材料,利用自然光气体熏蒸平台,设置对照(9 nL·L^-1)和高浓度臭氧(约100 nL·L^-1)处理,研究臭氧胁迫对稻穗不同部位糙米直链淀粉含量和RVA谱特征值的影响.结果表明: 与对照相比,臭氧处理使糙米直链淀粉含量、最高粘度、热浆粘度、崩解值和冷胶粘度分别下降5.9%、7.6%、5.9%、11.6%、2.9%,消减值和糊化温度分别增加24.9%和1.0%,均达显著水平.稻米直链淀粉含量和所有RVA特征参数的品种间差异均达极显著水平.稻穗不同部位稻米直链淀粉含量、最高粘度、热浆粘度、崩解值、冷胶粘度从大到小依次为强势粒>中势粒>弱势粒,消减值则表现相反.绝大多数情形下,臭氧与年度或臭氧与品种间的互作对稻米直链淀粉含量和RVA谱的影响达显著水平;尽管稻米RVA特征值对臭氧胁迫的响应多表现为稻穗上部略小于中部和下部,但臭氧与部位间的互作均未达显著水平.表明中等强度的臭氧胁迫使水稻食味品质明显变劣,变劣的程度因生长季和供试品种而异,籽粒着生位置受臭氧胁迫的影响较弱.     

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质。以农杆菌为介导进行转基因,获得9个转基因株系,从后代中筛到2个大片段缺失的变异株系L8和L9。它们的Wx基因表达大幅下调,直链淀粉含量极显著降低,胶稠度和食味品质也有所改善。L8和L9糙米外观有很大差异,L8与糯米相近,L9与粘米类似。综上所述,本研究利用CRISPR/Cas9技术编辑Wx基因编码区上游序列,获得两份低直链淀粉含量水稻种质,该结果为稻米品质改良研究提供了参考。     

用于筛选直链淀粉含量为中等的籼稻品种的分子标记

用PCR AccⅠ分子标记检测方法 ,检测了来自不同地区的 6 3个栽培水稻品种 (系 )蜡质基因第 1内含子剪接供体 1位碱基是G或是T。另外 ,还测定了这些水稻成熟种子的直链淀粉含量。结果显示该位置是G碱基的水稻品系成熟种子中直链淀粉含量均高于 2 0 % ,该位置是T的均低于 18%。在杂交育种过程中 ,这一分子标记可用于预测水稻植株种子的直链淀粉含量。对高直链淀粉含量的水稻亲本与中等直链淀粉含量的水稻亲本之间 5个籼型杂交组合F2 群体的分析表明 ,蜡质基因第 1内含子 1位碱基是G或是T与水稻种子中直链淀粉含量的高或低是紧密连锁 ,共同分离的。这些结果表明PCR AccⅠ分子标记检测方法可用于选育中等直链淀粉含量的籼稻新品系     

生育前期遮光对水稻后期功能叶生长及稻米品质的影响

以2个不同直链淀粉含量的水稻品种为材料,研究了生育前期(插秧至幼穗分化期)遮光对水稻后期功能叶生长及稻米品质的影响。结果表明,前期遮光处理后,水稻剑叶面积明显增大,功能叶的碳、氮代谢水平提高;2个品种的籽粒直链淀粉积累过程受到的影响不同,而蛋白质积累受到的影响均一致(即百分含量下降幅度增大);糙米率有所降低,但未达到显著水平;精米率和垩白率极显著下降;整精米率和直链淀粉含量显著或极显著升高;粒长、粒宽和蛋白质含量无显著性变化,但蛋白质含量表现为下降趋势;2个品种间存在着差异。     

Identification of optimal hyperspectral bands for estimation of rice biophysical parameters

The present study aims to identify the narrow spectral bands that are most suitable for characterizing rice biophysical parameters. The data used for this study come from ground-level hyperspectral reflectance measurements for five rice species at three levels of nitrogen fertilization during the growing period. Reflectance was measured in discrete narrow bands between 350 and 2 500 nm. Observed rice biophysical parameters included leaf area index （LAI）, wet biomass and dry biomass. The stepwise regression method was applied to identify the optimal bands for rice biophysical parameter estimation. This research indicated that combinations of four narrow bands in stepwise regression models explained 69% to 83% variability for LAI, 56% to 73% for aboveground wet biomass and 70% to 83% for leaf wet biomass. An overwhelming proportion of rice information was in a particular portion of near infrared （NIR） （1 100-1 150 nm）, red-edge （700-750 nm）, and a longer portion of green （550-600 nm）. These were followed by the moisture-sensitive NIR （950-1 000 nm）, the intermediate portion of shortwave infrared （SWlR） （1 650-1 700 nm）, and another portion of NIR （1 000-1 050 nm）.     

利用高光谱参数反演水稻叶片类胡萝卜素含量

为了探讨快速、准确预测水稻(Oryza sativa)叶片类胡萝卜素(Car)含量的敏感光谱波段和光谱指数, 通过实施涉及不同年份、不同生态点、不同施氮水平和不同品种类型的4个田间试验, 于主要生育期同步测定了水稻顶部4张叶片的光谱反射率及Car含量, 系统分析了350-2 500 nm范围内任意两波段组合而成的比值(SR (λ1, λ2))、归一化(ND (λ1, λ2))及已报道的对Car敏感的光谱指数与水稻叶片Car含量间的定量关系。结果表明, 不同Car含量水平下水稻叶片光谱反射率存在着明显变化, 以绿光及红边波段对水稻叶片Car含量变化最为敏感。723 nm附近的波段与近红外波段的比值组合以及713 nm附近的波段与近红外波段的归一化组合可以较好地预测水稻叶片Car含量, 以SR (723, 770)和ND (770, 713)表现最好, 线性拟合R²分别达到0.897和0.898。基于独立的试验资料的检验表明, 预测值和实测值的拟合R²分别为0.856和0.858, 均方根差RMSE均为0.072, 平均相对误差RE分别为11.9%和12.0%, 表明SR (723,770)和ND (770, 713)可有效地估算水稻上部叶片的Car含量。     

Mapping the myoglobin concentration, oxygenation, and optical pathlength in heart ex vivo using near-infrared imaging

A method that provides maps of absolute concentrations of oxygenated and deoxygenated myoglobin (Mb), its oxygenation, and its near-infrared (NIR) optical pathlength in cardiac tissue was developed. These parameters are available simultaneously. The method is based on NIR diffuse reflectance spectroscopic imaging and specific processing of the NIR images, which included a first derivative of the diffuse reflectance spectrum. Mb oxygenation, total Mb concentration, and NIR light pathlength were found to be in the range of 92%, 0.3 mM, and 12.5 mm, respectively, in beating isolated buffer-perfused and arrested pig hearts. The charge-coupled device camera enables sub-millimeter spatial resolution and spectroscopic imaging in 1.5 to 2.0 min. The technique is noninvasive and nondestructive. The equipment has no mechanical contact with the tissue of interest, leaving it undisturbed.     

Accuracy of near-infrared interactance instruments and population-specific equations for estimating body composition in young wrestlers

The purpose of this investigation was to determine the accuracy of near-infrared interactance (NIR) instruments and population-specific NIR equations for estimating percent body fat (% fat) in young wrestlers. Forty-four white male youth wrestlers (mean age +/- SD = 11.0 +/- 2.1 years) volunteered for this study. Thirteen NIR % fat estimates were cross-validated against the criterion % fat from underwater weighing. The results of this study indicated that the total error (TE) values associated with the NIR instrument generated % fat estimates as well as % fat estimates from population-specific NIR equations developed for high school wrestlers or adult men were too large (TE = 5.7-27.3% fat) to be used with young wrestlers. Based on the present findings, it is recommended that new NIR equations be developed and validated specifically for use in young male athletes.     

Dry matter production and boron concentrations of vegetative and reproductive tissues of canola and sunflower plants grown in nutrient solution

Prediction of nitrogen (N) mineralization is important for specifying the optimum rate of N fertilizer for flooded rice at the time of sowing. To develop a predictive test, soils (0–0.1 m) were sampled from 22 farms throughout the rice-growing region of southern Australia over a 4-year period. Near infrared reflectance (NIR) spectra of the soils were compared with sixteen biological and chemical soil tests for the prediction of N-uptake by rice plants from these soils in the field and glasshouse. The aim of the study was to develop a soil-NIR calibration as an accurate, rapid and economical mineralization test. Nitrogen uptake by field-grown and glasshouse-grown plants was poorly correlated (r = 0.30), even though significant NIR calibrations were developed with both. Since N uptake by rice in the field was affected by varying weather and management, the field calibration is probably spurious. The calibration of soil NIR spectra with N uptake by glasshouse plants was satisfactory, with a standard error (SE) of 13 kg ha^–1 over a range of 11 – 95 kg ha^–1, and a correlation between calculated and measured N uptake (r = 0.87, P<0.001). An even better soil-NIR calibration was found with N-mineralization after 21 days of anaerobic incubation (SE 16 mg kg^–1, range 52–175 mg kg^–1). Analysis of the soil spectra showed that similar wavelengths were correlated with both plant-N uptake and mineralization. NIR spectroscopy shows considerable potential to predict soil N mineralization, and may assist future fertiliser decision support.     

NIR techniques create added values for the pellet and biofuel industry

A 2(3)-factorial experiment was carried out in an industrial plant producing biofuel pellets with sawdust as feedstock. The aim was to use on-line near infrared (NIR) spectra from sawdust for real time predictions of moisture content, blends of sawdust and energy consumption of the pellet press. The factors varied were: drying temperature and wood powder dryness in binary blends of sawdust from Norway spruce and Scots pine. The main results were excellent NIR calibration models for on-line prediction of moisture content and binary blends of sawdust from the two species, but also for the novel finding that the consumption of electrical energy per unit pelletized biomass can be predicted by NIR reflectance spectra from sawdust entering the pellet press. This power consumption model, explaining 91.0% of the variation, indicated that NIR data contained information of the compression and friction properties of the biomass feedstock. The moisture content model was validated using a running NIR calibration model in the pellet plant. It is shown that the adjusted prediction error was 0.41% moisture content for grinded sawdust dried to ca. 6-12% moisture content. Further, although used drying temperatures influenced NIR spectra the models for drying temperature resulted in low prediction accuracy. The results show that on-line NIR can be used as an important tool in the monitoring and control of the pelletizing process and that the use of NIR technique in fuel pellet production has possibilities to better meet customer specifications, and therefore create added production values.     

Monitoring leaf photosynthesis with canopy spectral reflectance in rice

Non-destructive and rapid method for assessment of leaf photosynthetic characteristics is needed to support photosynthesis modelling and growth monitoring in crop plants. We determined the quantitative relationships between leaf photosynthetic characteristics and canopy spectral reflectance under different water supply and nitrogen application rates. The responses of reflectance at red radiation (wavelength 680 nm) to different water contents and nitrogen rates were parallel to those of leaf net photosynthetic rate (P _N). The relationships of reflectance at 680 nm and ratio index of R(810,680) (near infrared/red, NIR/R) to P _N of different leaf positions and leaf layers in rice indicated that the top two full leaves were the best leaf positions for quantitative monitoring of leaf P _N with remote sensing technique, and the ratio index R(810,680) was the best ratio index for evaluating leaf photosynthetic characteristics in rice. Testing of the models with independent data sets indicated that R(810,680) could well estimate P _N of top two leaves and canopy leaf photosynthetic potential in rice, with the root mean square error of 0.25, 0.16, and 4.38, respectively. Hence R(810,680) can be used to monitor leaf photosynthetic characteristics at different growth stages of rice under diverse growing conditions.     

Assessment of ATR‐NIR and ATR‐MIR spectroscopy as an analytical tool for the quantification of the total polyphenols in Dendrobium huoshanense

In this study, the potentiality of applying attenuated total reflectance near‐infrared (ATR‐NIR) and attenuated total reflectance mid‐infrared (ATR‐MIR) techniques combined with a partial least squares (PLS) regression technology to quantify the total polyphenols (TPs) in Dendrobium huoshanense (DHS) was investigated and compared. The real TP contents in the DHS samples were analysed using methods of reference. The capability of the two IR spectroscopic techniques to quantify the TPs in DHS was assessed by the root‐mean‐square error of calibration (RMSEC) and determination coefficients (R²). The results showed that both NIR and MIR might be used as a fast and simple tool to replace traditional chemical assays for the determination of the TP contents in DHS, and the best NIR model showed slightly better prediction performance [root‐mean‐square error of prediction (RMSEP): 0.307, R²: 0.9122, ratio performance deviation (RPD): 4.43] than the best MIR model (RMSEP: 0.440, R²: 0.9069, RPD: 3.09). Results from this study indicated that both the NIR and MIR models could be used to quantify the TP in DHS, and ATR‐NIR appeared to be the more predominant and more robust technique for the quantification of the TP in DHS.     

Direct prediction of bioethanol yield in sugar beet pulp using near infrared spectroscopy

Sugar beets are a raw material for the production of sugar and ethanol. The decision on which end product to pursue could be facilitated by fast and reliable means of predicting the potential ethanol yield from the beets. A Near Infrared (NIR) Spectroscopy-based approach was tested for the direct prediction of the potential bioethanol production from sugar beets. A modified partial least squares (MPLS) regression model was applied to 125 samples, ranging from 21.9 to 31.0 gL(-1) of bioethanol in sugar beet brei. The samples were analyzed in reflectance mode in a Direct Contact Food Analyser (DCFA) FOSS-NIRSystems 6500 monochromator, with standard error of cross validation (SECV), standard error of prediction (SEP), coefficient of determination (r(2)) and coefficient of variation (CV) of 0.51, 0.49, 0.91 and 1.9 gL(-1), respectively. The NIR technique allowed direct prediction of the ethanol yield from sugar beet brei (i.e. the product obtained after sawing beets with a proper machine) in less than 3 min.     

Plasmonic Ladder-Like Structure for NIR Polarizer

Polarization-dependent light transmission property is investigated in two-dimensional plasmonic ladder-like structure in the Near-infrared (NIR) regime of 900 to 1600 nm. The plasmonic ladder-like structures are fabricated using cost-effective laser interference lithography. Optical transmission studies reveal that in the stated NIR regime, the structure has nearly 30 % absolute transmission with respect to air when the long axis is aligned parallel to the polarization axis of the incident excitation and has negligible transmission at the crossed polarization state. The findings have potential implications in designing large area flat NIR polarizers.