Indication of the expression of transgene in rice plant based on hyperspectral remote sensing technique II--growth monitoring of samples in the contrast experiment.

Since the complication of monitoring and evaluating the problems about the transgenic expression and its impacts on the receptor in the transgenic crop breeding and other relevant evaluated works, the authors in the present work tried to assess the differences of spectral parameters of the transgenic rice in contrast with its parent group quantitatively and qualitatively, fulfilling the growth monitoring of the transgenic samples. The spectral parameters (spectral morphological characteristics and indices) chosen are highly related to internal or external stresses to the receipts, and thus could be applied as indicators of biophysical or biochemical processes changes of plant. By ASD portable field spectroradiometer with high-density probe, fine foliar spectra of 8 groups were obtained. By analyzing spectral angle and continuum removal, the spectral morphological differences and their locations of sample spectra were found which could be as auxiliary priori knowledge for quantitative analysis. By investigating spectral indices of the samples, the quantitative differences of spectra were revealed about foliar chlorophyll a+b and carotenoid content. In this study both the spectral differences between transgenic and parent groups and among transgenic groups were investigated. The results show that hyperspectral technique is promising and a helpful auxiliary tool in the study of monitoring the transgenic crop and other relevant researches. By this technique, quantitative and qualitative results of sample spectra could be provided as prior knowledge, as certain orientation, for laboratory professional advanced transgenic breeding study.

[Detection of the expression of transgene in rice plant based on hyperspectral remote sensing technique].

The present study aims to identify the expression of transgene in given rice plant samples in certain conditions. To avoid external noise caused by temperature change and water-loss, field spectrum was collected with ASD field spectrometer in natural state. The study calculated the mean spectrum of samples as main data set analyzed which were controlled by inner clustering coefficient to ensure data quality. By mean spectrum, the noise from random distinctions in few individual cultivators, which could not be expressed in the class stably, could be weakened even with filtering. With the help of parameters, such as red edge and green peak, this study gave qualitative spectral differences between transgenic samples and their parents. The results show that the transgenes in rice plant were expressed and influenced the samples. Moreover, it was found that the parameters of area are more suitable for describing the differences/changes of the samples, while PRI and SR-PRI are more sensitive to indicate them. Most of the above results could be found on the continuum-removal spectrum curve of samples. These conclusive results demonstrate that hyperspectral remote sensing technique has good prospects and application potential in transgene expression detection and monitoring, especially in plant breeding process.

[Quick testing-technology of transgenic rice with npt- screen marker gene].

By using npt-gene as assistant selection-marker, treating Japonica rice Zhonghua 9 [ZH9 (CK)] and transferred lysozyme gene rice (the donor rice is Japonica rice Zhonghua 9) [ZH9(R)] with antibiotics, we built a system of quickly testing transgenic rice offspring. Detached leaves of ZH9(R) and ZH9(CK) were treated by Kanamycin (0, 400, 450, 500, 550, 600 and 700 mg/L) and G418 (0, 40, 60, 80, 100, 150 and 200 mg/L) with different concentrations. The result show effect of Kanamycin is not evident and G418 is the best antibiotics to test transgenic rice with npt- gene. The data showed that 80 mg/L G418 (treating 4 d) was optimal to test transgenic rice. Further study was done with seeds, young embryos and seedlings by G418 testing: the alive seeds were cultured in the culture dishes which filled with a series of G418 solution with different concentration of 0, 100, 150, 200, 250, 300 and 350 mg/L; in the tissue culture room, the germinating embryos were inoculated in the culture medium (1/2 MS+0.5 mg/L 6-BA+1.5% sucrose) which contains G418 with 0, 150, 200, 250 and 300 mg/L concentration; the aseptic seedlings were inoculated in the the same culture medium (1/2 MS+0.5 mg/L 6-BA+1.5% sucrose) which contains G418 with 0, 100, 150, 200 and 250 mg/L concentration. The conclusions indicated that 300 mg/L (treating 7 d) was the critical concentration to test seeds of transgenic rice; 200 mg/L (treating 10 d) was the critical concentration to test young embryo of transgenic rice; 150 mg/L (treating 12 d) was the critical concentration to test seedlings of transgenic rice. Two primers were designed based on npt- and lysozyme gene sequences. PCR technology confirmed the above detection system. The preliminary results showed npt-was tightly linked with lysozyme gene. Above confirmed critical concentrations were applied to test detached leaves, seeds, young embryos and seedlings of transferred generations. The effect was very obvious. It is convenient, intuitionistic, and exact way that aparting the positive plants from the mixture of transgenic positive and negative plants with G418.