Grain yield,nitrogen use efficiency and physiological performance of indica/japonica hybrid rice in response to various nitrogen rates

Utilizing the heterosis of indica/japonica hybrid rice(IJHR)is an effective way to further increase rice grain yield.Rational application of nitrogen(N)fertilizer plays a very important role in using the heterosis of IJHR to achieve its great yield potential.However,the responses of the grain yield and N utilization of IJHR to N application rates and the underlying physiological mechanism remain elusive.The purpose of this study was to clarify these issues.Three rice cultivars currently used in rice production,an IJHR cultivar Yongyou 2640(YY2640),a japonica cultivar Lianjing 7(LJ-7)and an indica cultivar Yangdao 6(YD-6),were grown in the field with six N rates(0,100,200,300,400,and 500 kg ha^(-1))in 2018 and 2019.The results showed that with the increase in N application rates,the grain yield of each test cultivar increased at first and then decreased,and the highest grain yield was at the N rate of 400 kg ha^(-1)for YY2640,with a grain yield of 13.4 t ha^(-1),and at 300 kg ha^(-1)for LJ-7 and YD-6,with grain yields of 9.4–10.6 t ha^(-1).The grain yield and N use efficiency(NUE)of YY2640 were higher than those of LJ-7 or YD-6 at the same N rate,especially at the higher N rates.When compared with LJ-7 or YD-6,YY2640 exhibited better physiological traits,including greater root oxidation activity and leaf photosynthetic rate,higher cytokinin content in the roots and leaves,and more remobilization of assimilates from the stem to the grain during grain filling.The results suggest that IJHR could attain both higher grain yield and higher NUE than inbred rice at either low or high N application rates.Improved shoot and root traits of the IJHR contribute to its higher grain yield and NUE,and a higher content of cytokinins in the IJHR plants plays a vital role in their responses to N application rates and also benefits other physiological processes.

Performance of Five Early Indica Rice Varieties under Direct Seeding in Jiangbei District

Comparative experiments were conducted on five conventional indica rice varieties,including Yongxian 15,Zhongzao 39,Shunda 135,Zhongzu 143,and Zhongzu 18.Under direct seeding conditions,their growth period and yield performance were investigated.The results showed that the yield of Zhonggang 143 and Shunda 135 was higher than that of Yongxian 15(CK),and Zhonggang 143 had the highest yield of 7.494 t/hm 2,followed by Shunda 135(7.467 t/hm 2);Yongxian 18(CK)has a yield of 7.326 t/hm 2.Taking into account factors such as field growth,yield,and growth period,Zhongzu 143 and Shunda 135 can be further promoted to optimize the early rice variety structure in Jiangbei District.

Developmental Genetic Analysis of Brown Rice Weight Under Different Environmental Conditions in indica Rice

Analysis of genetic main effects and genotype x environment (GE) interaction effects for brown rice weight (BRW) at four different filling stages in indica lice (Oryza sativa L.) was conducted for two-year experimental data by using developmental genetic models and corresponding statistical approaches for quantitative traits of seeds in cereal crops. It was indicated that the genetic main effects and their GE interaction effects of triploid endosperm, cytoplasmic and diploid maternal plant genes were important for BRW at different filling stages of rice, especially for endosperm or maternal additive main effects and their additive interaction effects. Because of the higher additive effects and additive interaction effects for BRW at different filling stages, the better improving effects for this trait could be expected by selection in rice breeding. The results of conditional genetic variance components showed that the new expression of quantitative genes in endosperm and maternal plant for BRW was mostly found at all different filling stages of rice. The gene expression, however, was most active at the early filling stages especially for the first (1-7 d) and the second filling stages (8-14 d after flowering). The phenomena that some genes were spasmodically expressible among filling stages of rice were detected for some genetic effects especially for net cytoplasmic main effects or its interaction effects and net dominance main effects. Predicted genetic effects at different filling stages of rice showed that some parents such as V20 and Zuo 5 were better than others for improving the BRW.

Analysis of the Conditional Correlations from Different Genetic Systems Between the Protein Content and the Appearance Quality Traits of Indica Rice

A factorial mating design in two environments was conducted using 7 cytoplasmic male sterile lines （A） and 5 restorer lines （R） along with their F1 （A × R） and F2 populations. The unconditional and conditional analyses of genetic models and the corresponding statistic methods, including endospermic, cytoplasmic, and maternal plant genetic systems, were used to analyze the genetic relationships between protein content （PC） and the appearance quality traits of indica rice （Oryza sativa L.）. The results from unconditional analysis indicated that PC was significantly correlated with the appearance quality traits of rice, except for the brown rice thickness （BRT）. Only the genetic covariance between PC and the brown rice width （BRW） was positively correlative, whereas all the other pairwise traits were negatively correlative. The results from conditional analysis revealed that the weight of brown rice （WBR） or the amylose content （AC） could significantly affect the relationships between PC and the appearance quality traits of indica rice. The conditional analysis showed that WBR might negatively affect the relationships between PC and the brown rice length （BRL）, BRW, or BRT through the geuotype x environmental （GE） interaction effects, but positively affected the relationships between PC and the ratio of brown rice length to width （RLW） or the ratio of brown rice length to thickness （RLT）. The amylase content could positively affect the relationships between PC and BRL, RLW, RLT through the cytoplasmic effects and maternal additive effects, but negatively affected the relationships between PC and BRW.

Development of Isogenic Lines of Morphological Markers in Indica Rice

A total of 28 accessions of rice (Oryza sativa L.) germplasms were collected from International Rice Research Institute (IRRI), Philippine and Gene Pool of China National Rice Research Institute (CNRRI). These germplasms were attributed as a Donor Gene Pool, consisting of 27 diverse morphological marker lines with each having a unique valuable trait (gene), and the recurrent parent-Zhefu 802, an early-season indica cultivar in south of China in the 1980s and 1990s. A set of 27 isogenic lines with 2-3 markers on every chromosome were obtained after more than 10 successive backcrosses. Moreover, 15 accessions of double-marker isogenic lines were developed by further pyramiding markers on the same chromosome. Key agronomic traits such as heading date, plant height, tillering number and panicle length in this set of lines were similar to that of Zhefu 802 except the marker traits.

Correlation Analysis of Characters of Indica Hybrid Rice with High Content of Amylose

[Objective] The aim was to research and analyze characters and related correlation of indica hybrid rice containing high amylose. [Method] In the research, major characters of 16 species of groups of {ndica hybrid rice and the correlation were explored in rice belts in Sichuan in 2011. [Result] The variation of chalkiness degree was the highest at 26.8%, followed by rate of head rice at 22.89%; varia- tions of head rice rate and chalkiness degree were lower at 0.72% and 2.61%. In addition, rate of unpolished rice was of significantly negative correlation with chalki- ness degree; rate of chalky rice was of extremely positive correlation with chalkiness degree and gel consistency; chalkiness degree was of significantly positive correla- tion with gel consistency; gel consistency was of significantly negative correlation with protein content; amylose content was of insignificant correlation with gel consis- tency （r=0.166 9） of the 16 hybrid groups. These indicated that gel consistency would be still high as the content of amylose grows （AC≥25%）. For example, when the content of anylose in Chuanxiang 29A/Luhui 37, Kangfeng AJ6684, and Lefeng A/SR536 exceeded 27%, the gel consistency was over 80 mm in the research. [Conclusion] The research provides references for improvement of amylose content in breeding hybrid rice.

Difference of Nitrogen Uptake and Use Efficiency in Conventional Indica Rice Cultivars with Different Nitrogen Use Efficiency for Grain Output

[Objective] The study aimed to confirm difference of nitrogen uptake and used efficiency with different nitrogen use efficiency for grain output (NUEg) types of indica rice.[Method] 88 and 122 conventional indica rice cultivars were solution-cultured in 2001 and 2002, respectively. Dry matter weight (including root system, culm and sheath, leaves, panicle), nitrogen content of different organs, yield and its components were measured. The tested rice cultivars were classified into 6 types (i.e. A, B, C, D, E and F, A was the lowest, and F was the highest) based on their NUEg level by the MinSSw method.[Result](1)Difference of NUEg of the cultivars used in this study were very large; (2) No significant difference of N content at heading stage was observed among different NUEg types of indica rice. In the cultivars with higher NUEg, however, N content in leaf, stem-sheath and entire rice plant were lower at mature stage. (3)Cultivars with higher NUEg were characterized with lower N uptake before heading and at mature stage; (4) Cultivars with higher NUEg were characterized with higher N use efficiency in biomass production and harvest index. [Conclusion] The cultivars with higher NUEg showed lower N uptake and N content, while nitrogen use efficiency was higher.

Changes in Agronomic Traits of Indica Hybrid Rice during Genetic Improvement

[Objective]The aim was to elucidate the changes of agronomical traits of indica hybrid rice during its genetic improvement.[Method]23 typical indica hybrid combinations cultivated in the middle and lower reaches of Yangtze River during the last 30 years were selected as experimental materials.All the hybrid rice combinations were grown in the same field;and then yield and morphological characters of single plant were compared;correlation and path analyses among agronomic characteristics were also conducted.[Result]During the 30 years of genetic improvement,the effective panicle number per plant was progressively decreased,which caused the decrease of grain yield and biomass per plant of hybrid rice combinations in spite of the slightly increased plant height,1 000-grain weight and number of grains per panicle.Although the transformation percentage of the matter in stem and sheath decreased after flowering,leave photosynthetic capacity after flowering was increased as panicle weight.Correlation and path analysis showed that although both biomass per plant and number of grains per panicle were significantly correlated with grain yield per plant,biomass per plant acted as the key factor affecting yield per plant.[Conclusion]It was suggested that in rice high yield cultivation,panicle number was as important as large panicle,and tillering ability should not be neglected in breeding.

Breeding and Application of WA-type Indica CMS Line Ningxian 2A in Rice

Ningxian 2A is a new Indica CMS line with WA type sterile cytoplasm in rice. It was developed from the testcross between WA type indica CMS line Jin 23A and the single plant selected from the F_4 population of the cross Wuxiang 2B×Jin 23B followed by successive backcross over years. In September, 2014, it was technically identified by Jiangsu Provincial Crop Variety Appraisal Committee. Ningxian 2A has the characteristics of complete pollen sterility, high outcrossing rate, good plant type, high combining ability and good grain quality. Among its hybrids, Ningxian you 8（ Ningxian 2A×R08） was registered and released for commercial production in Jiangsu Province in May, 2015.

Analysis of Genetic Variation and Population Structure of Starch Synthesis-related Genes in Indica Rice Cultivars

In this study, 34 molecular markers of starch synthesis-related genes were used to evaluate the genetic variation and population structure of 87 indica rice cultivars from different countries and regions. The results showed that a total of 80 alleles were amplified using 34 primer pairs, with an average of 2.5 alleles per locus. The allele number varied from 2 to 6 among various cultivars. Shannon＇s diversity index of molecular markers varied from 0.303 to 0.796, with an average of 0.539. Polymorphism information content （PIC） varied from 0.084 to 0.658, with an average of 0.295. The genetic similarity coefficients of 87 indica rice cultivars ranged from 0.265 to 0.990, indicating significant genetic differences of starch synthesis-related genes among different cultivars, but the variation frequency of alleles varied among different cultivars. Population structure analysis showed that these 87 indica rice cultivars were divided into three categories. Genetic differences were small within the same category but great among different categories. Moreover, indica rice cultivars with simple genetic components accounted for 39.1% and those with complex genetic background accounted for 60.9%. This study may not only provide theoretical basis for genetic improvement of rice starch quality, but also lay a solid foundation for subsequent association analysis of rice quality-related traits.

Analysis of Heterosis, Combining Ability and Heritability of Cadmium Content in Brown Rice of Three-line Indica Hybrid Rice

Five cytoplasmic male sterile （CMS） lines were used as parents in an incomplete diallet cross and six restorer lines of rice design （Nc II design）. Thirty hybrid combinations with the same growth period were selected as experimental ma- terials to investigate the heterosis, combined ability and heredity of Cd content in brown rice of indica hybrid rice. According to the results, Cd content in brown rice showed a significantly negative heterosis; the general combining ability and specific combination ability of Cd content in CMS and restorer lines both reached extremely significant level （P〈0.01）, indicating that both genetic improvement of parents and e- valuation of combinations are important to the breeding of hybrid combinations with low accumulation of Cd; the broad-sense heritabitity and narrow-sense heritability of Cd content were both relatively high with slight differences, which respectively reached 97.73% and 80.10%, indicating that Cd content in brown rice mainly de- pends on the additive action of genes; in addition, parent improvement showed bet- ter effect on the selection of early generation.

Synthesis and paste properties of octenyl succinic anhydride modified early Indica rice starch

Octenyl succinic anhydride （OSA） modified early Indica rice starch was prepared in aqueous slurry systems using response surface methodology. The paste properties of the OSA starch were also investigated. Results indicated that the suitable parameters for the preparation of OSA starch from early Indica rice starch were as follows： reaction period 4 h, reaction temperature 33.4℃, pH of reaction system 8.4, concentration of starch slurry 36.8% （in proportion to water, w/w）, amount of OSA 3% （in proportion to starch, w/w）. The degree of substitution was 0.0188 and the reaction efficiency was 81.0%. The results of paste properties showed that with increased OSA modification, the starch derivatives had higher paste clarity, decreased retrogradation and better freeze-thaw stability.

Quantitative Trait Loci for Grain Chalkiness and Endosperm Transparency Detected in Three Recombinant Inbred Line Populations of Indica Rice

Quantitative trait loci（QTL） for percentage of chalky grain,degree of chalkiness,and endosperm transparency were detected using 3 recombinant inbred line populations derived from crosses between parental lines of commercial three-line hybrids of indica rice.Two of the populations showed great variations on heading date,and the other had a short range of heading date variation.A total of 40 QTLs were detected and fell into 15 regions of 10 chromosomes,of which 5 regions were detected for 1 or more same traits over different populations,2 were detected for different traits in different populations,3 were detected for 2 or all the 3 traits in a single population,and 5 were detected for a single trait in a single population.Most of these QTLs have been reported previously,but a region located on the long arm of chromosome 10 showing significant effects in all the 3 populations has not been reported before.It was shown that a number of gene cloned,including the Wx and Alk for the physiochemical property of rice grain,and GW2,GS3 and GW5 for grain weight and grain size,could have played important roles for the genetic control of grain chalkiness in rice,but there are many more QTLs exerting stable effects for rice chalkiness over different genetic backgrounds.It is worth paying more attentions to these regions which harbor QTL such as the qPCG5.2/qDC5.2/qET5.2 and qPCG10/qDC10/qET10 detected in our study.Our results also showed that the use of segregating populations having high-uniform heading date could greatly increase the efficiency of the identification of QTL responsible for traits that are subjected to great environmental influence.

Natural Variation of Leaf Thickness and Its Association to Yield Traits in indica Rice

Leaf thickness is an important morphological trait in rice. Its association to the yield potential, as of now has not been documented because of the shortage of the equipment which could conveniently measure the leaf thickness in rice. In this study, the thickness of top three leaves of 208 cultivars had been determined by a nondestructive rice leaf thickness instrument for the research of the natural variation of leaves thickness and its association to yield traits in indica rice. The results showed that the flag leaf was the thickest, and the 2nd leaf was thicker than the 3rd leaf. Analysis of variance indicated the existence of wide genetic diversity of leaf thickness among the investigated indica rice genotypes. The tight correlation among the thicknesses of the top three leaves means that the leaf thickness traits share one genetic control system. Leaf thickness had a significant positive correlation with leaf length and a positive correlation with leaf width, indicated that thicker leaf was beneficial to increasing the single leaf area. The results of correlation analysis revealed that thicker leaf should be profitable to the leaf erection, higher numbers of grains per panicle and higher grains weight per panicle. However, the significantly negative correlation between leaf thickness and number of panicles per plant counteracted the profitability from increased grains weight per panicle, so that the correlations of the thicknesses of the top three leaves to yield and biomass were positive but not significantly. It has made great progress in the genetic improvement of leaves thickness in inbred indica rice breeding in Guangdong Province, China, since the 1990s.

Suitable growing zone and yield potential for late-maturity type of Yongyou japonica/indica hybrid rice in the lower reaches of Yangtze River, China

Late-maturity type of Yongyou japonica/indica hybrids series （LMYS） have shown great yield potential, and are being widely planted in the lower reaches of Yangtze River, China. Knowledge about suitable growing zone and evaluation of yield advantage is of practicall importance for LMYS in this region. Fifteen LMYS, two high-yielding inbred japonica check varieties （CK-J） and two high-yielding hybrid indica check varieties （CK-I） were grown at Xinghua （119.57°E, 33.05°N） of Lixiahe region, Yangzhou （119.25°E, 32.30°N）of Yanjiang region, Changshu （120.46°E, 31.41°N）of Taihu Lake region, and Ningbo （121.31°E, 29.45°N） of Ningshao Plain in 2013 and 2014. The results showed that maturity dates of the 15 were later than the secure maturity date at Xinghua and 6, 14 and 15 LMYS were mature before the secure maturity date at Yangzhou, Changshu and Ningbo, respectively. One variety was identified as high-yielding variety among LMYS （HYYS） at Yangzhou, 8 HYYS in 201：3 and 9 HYYS in 2014 at Changshu, 9 HYYS at Ningbo. HYYS here referred to the variety among LMYS that was mature before the secure maturity date and had at least 8% higher grain yield than both CK-J and CK-I at each experimental site. Grain yield of HYYS at each experimental site was about 12.0 t ha-1 or higher, and was significantly higher than CK varieties. High yield of HYYS was mainly attributed to larger sink size due to more spikelets per panicle. Plant height of HYYS was about 140 cm, and was significantly higher than check varieties. Significant positive correlations were recorded between duration from heading to maturity stage and grain yield, and also between whole growth periods and grain yield. HYYS had obvious advantage over check varieties in biomass accumulation and leaf area duration from heading to maturity stage. Comprehensive consideration about safe matudty and yield performance of LMYS at each experimental site, Taihu Lake region （representative site Changshu） and Ningshao Plain （representative site Ningbo） were thought suitable growing zones for LMYS in the lower reaches of Yangtze River. The main factors underlying high yield ofHYYS were larger sink size, higher plant height, longer duration from heading to maturity stage and whole growth periods, and higher biomass accumulation and leaf area duration during grain filling stage.

Developmental Genetics Analysis for Plant Height in indica Hybrid Rice Across Environments

The developmental genetics of plant height was analyzed from two groups of three-line indica hybrid rice at two environmental conditions based on the NCII design, using the additive-dominant developmental genetics models and the statistic methods. The results showed that the rice genotypes and environmental conditions could both affect plant height, and the effects of environment on plant height decreased gradually with plant development. Additive and dominant effects both governed the performance of plant height at all developmental stages. However, the degrees of effect varied among the rice genotypes. Moreover, the interaction between environments and genotypes also affected plant height. The genetic effects differed at most developmental stages. Furthermore, the expressJon of additive effect was more active than that of dominant effect. Conditional interaction effects with environment also influenced plant height during genetic development, especially at the eady stage. Mid-parent heterosis （HMP） increased gradually with the developmental stage of plant height, and maximized at the latest stage, whereas the heterosis over the better parent （HBP） showed small differences among the genotypes, and kept stable at the later stage, with positive numeric value. At most developmental stages, conditional HMP was positively significant, while conditional HBP was negatively significant. All above results suggest that HMP and HBP have some new expressions in all developmental periods and the levels and directions are quite different.

Effects of different mechanical direct seeding methods on grain yield and lodging resistance of early indica rice in South China

Direct seeding of rice has become a main planting method due to the low labor input and high economic benefit in South China.Dry direct seeding(DDS)has been widely used for single-season rice planting establishment.However,few studies have examined the performance of early-season indica rice under mechanical dry direct seeding.A two-year field experiment was conducted with two indica rice cultivars(i.e.,Zhongjiazao 17 and Zhuliangyou 819)to study lodging characteristics and grain yield formation under DDS,flooded direct seeding(FDS)and wet direct seeding(WDS)patterns.The results showed that the annual grain yield in DDS was higher by 14.42–26.34%for cultivar ZLY819 and 6.64–24.58%for cultivar ZJZ17 than in WDS and FDS,respectively,and these increases were mainly attributed to the improvement of the panicles.The DDS pattern significantly increased the seedling emergence rate of early indica rice cultivars,and increased total dry weight and crop growth rate.Meanwhile,shorter basal internodes,better stem diameter and stem wall thickness and lower lodging index were found in DDS in contrast to FDS and WDS.In particular,DDS improved the stem lodging resistance.Our results suggested that the appropriate direct seeding method was beneficial for improving the grain yield and lodging resistance of early indica rice.

Response of grain-filling rate and grain quality of mid-season indica rice to nitrogen application

Nitrogen is one of the important factors for high yield of rice.Apart from high yield,high quality has become the current urgent demand for rice production.Grain-filling stage is crucial for rice yield and quality formation.However,the effect of nitrogen on grain-filling characteristics and the relationship of grain-filling characteristics and rice quality of mid-season indica rice were still unclear.A field experiment was carried out to ascertain the critical grain-filling characteristics that contribute to rice milling quality,appearance quality and cooking and eating quality under nitrogen applications.The results showed that nitrogen applications prolonged the duration of superior and inferior grain filling.The mean grainfilling rate(G_(mean))and the maximum grain-filling rate(G_(max))of the inferior grains were positively correlated with chalky kernel rate,chalkiness,and amylose content.The time reaching the maximum grain-filling rate(T_(max)G)of the inferior grains was positively correlated with brown rice rate,milled rice rate,and head milled rice rate.Chalky kernel rate and chalkiness were negatively correlated with peak paste viscosity and breakdown viscosity.Less amylose content and more crude protein content were detected in nitrogen application of Liangyoupei 9 and Y Liangyou 2 both in 2016 and 2017.According to the correlation analysis,better cooking and eating quality of Y Liangyou 2 which had less amylose content might result from its higher G_(max)and G_(mean)of inferior grain than that of Liangyoupei 9 in the treatments of nitrogen application.These results indicated that the prolonging grain-filling duration and increasing grain weight at the maximum grain-filling rate of inferior grains contributed to the improvement of milling quality,appearance quality,and cooking and eating quality of mid-season indica rice under appropriate nitrogen applications.

Effects of increasing panicle-stage N on yield and N use efficiency of indica rice and its relationship with soil fertility

Because of the higher nitrogen(N)recovery efficiency(NRE)of panicle-stage fertilization compared with basal and tillering fertilization,increasing the proportion of N topdressing at the booting stage(panicleN)is recommended and commonly practiced in parts of China.To investigate the effects of increasing panicle-N on grain yield and N use efficiency(NUE)and the relationships of the increase and the rice cultivar and soil fertility status,we increased the percentage of panicle-N from 20%to 40%by correspondingly reducing the N amount applied only at the tillering stage in both high-and low-fertility blue clayey paddy fields in 2018 and 2019.Four indica cultivars with diverse panicle types were used,and their grain yield,dry matter accumulation,and NUE were compared.In high-fertility soil,increasing topdressing panicle-N from 20%to 40%reduced tillering ability and reduced the effective panicle numbers of the multi-and medium-panicle cultivars Huanghuazhan(HHZ),C Liangyouhuazhan(CHZ),and Tianyouhuazhan(THZ).These cultivars gave the greatest yield when 30%of N was supplied as panicle fertilizer,whereas the yield,NRE,N agronomic efficiency(NAE),and nitrogen physiological efficiency(NPE)of the heavy-panicle inbred cultivar Yangdao 6(YD6)continued to increase,resulting in improved dry matter accumulation and grain filling in the late growth stage.The yield,NAE,NRE,and NPE of YD6 peaked when the panicle-N constituted 40%.While in low-fertility soil,the multipanicle cultivar HHZ showed the greatest yield when 30%of fertilizer-N was applied once at the panicle initiation(PI)stage,while the medium-panicle cultivar CHZ showed the greatest yield when the panicle-N percentage was 40%.Our results suggest that the percentage of panicle-N fertilizer should not exceed 30%for multipanicle cultivars,while can be appropriately increased to 40%for heavy-panicle indica cultivars.The effect of increasing topdressing panicle-N on the yield of medium-panicle cultivars was related to soil fertility.The optimum panicle-N percentage was 30%in the high-fertility soil and 40%in the low-fertility soil.

Optimization of Electroporation Parameters for Immature Embryos of indica Rice (Oryza sativa)

To obtain a suitable condition for electroporation transformation in indica rice, the 10-day-old immature embryos were selected for optimization experiments. The results showed that one pulse at 850 V/cm, 950μF capacitance, 200 μL electroporation buffer with 70 mmol/L sodium glutamate, 100 μg/mL plasmid, 50μg/mL carrier DNA, 20 embryos per cuvette, 0℃ treatment and CC medium were the best parameters, which not only improved the transformation efficiency to 30.89%, but also ameliorated the embryo survival ratio to 95.92%. To further verify the practicability of this condition, the embryos from another indica rice variety and a rice type Ⅱ metallothionein-like gene （OsMT2bL） promotec：mgfp5：：gusA construct were tested, and specific GUS expression on the embryos was visualized by histochemical staining. The results showed that the GUS expression on the embryos activated by the OsMT2bL promoter was mainly concentrated on the apical point of the plumule whereas the expression driven by CaMV35S promoter was distributed on nearly all areas of the electroporated tissues. These results indicated that the optimized embryo electroporation conditions could be used not only in genetic transformation of indica rice but also in assay of gene regulation on embryos.