出苗难是贵州省酒用高粱产业的一个"瓶颈",本研究旨在探索适宜的酒用高粱出苗提升技术.以当地主栽高粱品种"红缨子"为试材,设置种子处理、土壤类型和播种深度的三因素实验,研究各因素及其互作对高粱出苗的影响.在人工气候箱中,CG包衣种子发芽率、芽长、发芽指数、平均发芽日数和活力指数与对照均无显著差异(p<0.05).在盆栽条件下,CZ和CG包衣种子在泥炭土、黄壤和石灰土中 3 cm,5 cm和 7cm播种深度均表现出较高的出苗率(p<0.05),而P2 丸化种子在石灰土中 3cm和 7cm播种深度表现出较高的出苗率(p<0.05);多因素方差分析表明,种子处理和土壤类型互作影响出苗率(p<0.05).在大田条件下,各试点结果均表明,CZ 和 CG 包衣种子及P2丸化种子出苗率显著高于对照(p<0.05).研究表明,种子经CG包衣处理,在 3cm播种深度下出苗率明显提高,值得今后推广.

A survey was conducted on the soils in the concentrated distribution area of ancient tea trees in Liangyan Town of Guizhou Province, and the nutrient contents and correlation among nutrients of 31 soil samples were analyzed, and the soil nutrient status was comprehensively evaluated by calculating the integrated index of soil fertility(IFI) using principal component analysis. The results showed that the average pH value of ancient tea plantations in Liang Yan was 5.37, and 51.61% of the soils were in the pH range of 4.5–5.5, which was generally suitable; the soil organic matter and alkaline nitrogen content were rich; however, the soil fast-acting phosphorus content did not reach the fertility index of high-yielding and high-quality tea plantations, and was in the state of phosphorus deficiency; the contents of exchangeable aluminum, exchangeable calcium and effective copper in most of the soils were in the suitable range for the growth of tea trees; the content of available iron and available zinc in the soil of ancient tea forest is low, with only 48.38% of the effective zinc in the suitable range. The principal component analysis showed that the soil fertility of ancient tea forest in Sanlitian Village was the best, followed by Guangou Village, and the soil standing conditions of ancient tea plant sites in Feilun, Taiji and Liangyan Villages were poor, and the focus should be on increasing phosphorus fertilizer and increasing iron, zinc and other trace elements appropriately for the poor fertility areas.

水稻穗部性状与产量直接相关,筛选控制水稻穗部性状的基因对于水稻育种具有重要意义.本研究以日本晴为母本,染色体片段代换系Z1347为父本构建F2群体,用SSR标记对穗长、二次枝梗数等穗部性状进行了QTL定位.结果表明:Z1347的穗长和二次枝梗数均显著高于日本晴,但两者的一次枝梗数无差异;SSR标记检测出5个QTL,其分布于水稻第1、6、7和11染色体上,其中与穗长相关的qPL1、qPL6、qPL7和qPL11等4个QTL,分别与RM5389、RM2126、RM6063和RM457连锁,与二次枝梗数相关的QTL(qNSB7)与RM6063连锁.5个QTL中,qPL11和qNSB7可能为新的位点,可在后续试验中加强其遗传效应的研究,为进一步的分子辅助育种提供参考.

[目的]明确新不育系配合力及遗传参数,为其优势组合选配提供依据.[方法]选用5个新选三系不育系和10个恢复系,按不完全双列杂交设计组配50个杂交组合,分析株高、有效穗数、穗长、每穗实粒数、每穗总粒数、结实率、千粒重和单株产量等8个农艺性状配合力,并估算其遗传参数.[结果]株高、有效穗数、每穗实粒数、结实率和单株产量等5个性状的一般配合力方差均达极显著差异水平(P<0.01,下同),说明这些性状的遗传主要受加性基因控制,穗长、每穗总粒数和千粒重等3个性状的一般配合力和特殊配合力方差均达显著(P<0.05)或极显著差异水平,说明这些性状的遗传受加性和非加性基因共同控制.118A的株高、有效穗数、结实率、千粒重和单株产量等5个性状的一般配合力效应值最高,134A的穗长和每穗实粒数的一般配合力效应值最高,141A的每穗总粒数的一般配合力效应值最高,不同不育系对其配制的F1代性状表型影响存在明显差异.单株产量特殊配合力效应值较高的组合是118A×SR498、141A×蜀恢527和134A×华占,均在0.0060以上.不育系在有效穗数、每穗实粒数、每穗总粒数和单株产量等4个性状的基因型方差明显高于恢复系,其对F1代的该性状表现起主要作用.[结论]118A、134A和914A是较理想的不育系材料,具有较好的应用前景;118A×SR498、141A×蜀恢527和134A×华占为苗头组合,可参加预试.

对水稻染色体片段代换系Z745进行染色体代换片段鉴定,并用以日本晴为背景亲本、水稻染色体片段代换系Z745为供体亲本杂交构建的次级F2群体定位水稻穗部性状QTL,以期发现新的穗部性状QTL.结果发现,Z745共含有17个代换片段,分布于第1、3—8、10和12染色体,代换片段总长度为26.1 Mb,平均代换长度为1.54 Mb;Z745的穗长、一次枝梗数、二次枝梗数和穗粒数均极显著高于日本晴;鉴定出21个穗部性状QTL,分布于其中9个代换片段上,位于第1、3—5、7—8、10和12共8条染色体上,其中,穗长QTL 4个(1、4和7染色体)、一次枝梗数QTL 4个(1、5、7和8染色体)、二次枝梗数QTL 6个(1、7、10和12染色体)、穗粒数QTL 7个(1、3、7、8、10染色体).定位到的穗长QTL(qPL7-1、qPL7-2)、二次枝梗数QTL(qNSB7-1)和穗粒数QTL(qSPP1-1)已被报道,其余穗部性状QTL尚未报道.

[Objective]The present study aimed to map and analyze narrow leaf mutant gene in order to provide the ba-sis for leaf developmental regulation and plant-type breeding of rice. [Method]Luhui17 was induced by ethyl methyl sul-fonate(EMS) and then a stable narrow leaf mutant(Narrow leaf 11,nal11) was obtained. Length and width of functional leaves, tiller number and plant height at maturation stage of nal11 were analysed and compared with those of wild type Mi-anhui 727. Orthogonal and reciprocal crosses between nal11 and Mianhui 727 were carried out and F2 population was ob-tained, and genetic analyzis and gene mapping were conducted. [Result]Widths of flag leaf and the 2nd and 3rd leaves from the top of nal11 at heading stage were significantly(P<0.05,the same below) or extremely significantly(P<0.01,the same below)different from those of wild type Mianhui 727, and were 60.7%, 57.9%, and 75.8% of the value for wild-type Mianhui 727 respectively. However, there was no difference in length of functional leaves between them(P>0.05). Plant height of nal11 was 90.3% of that of wild type Mianhui 727 and there was significant difference between them. Tiller num-ber of nal11 extremely significantly increased and was 150.0% of that of wild type Mianhui 727. In F1 generation of recip-rocal and orthogonal crosses between nal11 and Mianhui 727, the leaf width was normal in F1, but trait segregation occurred in leaf width of F2 populations. The ratio of rice plants with normal and narrow leaves was conformed as 3:1 by χ2 test. The results indicated that nal11 was recessive mutation controlled by nuclear gene. Using simple sequence repeat (SSR) marker, nal11 gene was located in a range of about 322 kb between RM7290 and RM16720 on chromosome 4 of rice. The genetic distances between nal11 and the two SSR markers were both 0.8 cM, covering 32.236 kb physical do-main. There were five annotation genes(Os04g26834,Os04g26850,Os04g26870,Os04g26880 and Os04g26841) in the region. The sequences were different from those of narrow leaf genes cloned by previous researchers. [Conclusion ]A new narrow leaf mutant (nal11) is obtained. The narrow leaf trait is controled by one recessive nuclear gene. It is located between RM7290 and RM16720 on chromosome 4 of rice, and has obvious influences on functional leaf, plant height and tiller number.

Genetic analysis of narrow leaf mutant may provide a reference for gene mapping and plant-shape breeding in rice.A narrow leaf mutant (nal 11) was induced by ethyl methyl sulfonate from Luhui 17.nal 11 was crossed reciprocally with Mianhui 727,R30 and Fuhui 838,respectively,to construct F2 populations.The length and width of the three functional leaves of nal 11 and Mianhui 727 were investigated at heading stage and the genetic analysis of nal 11 was also carried out.The results showed that:Length of the flag leaf of nal 11 was significantly shorter than cross-parent of Mianhui 727;length of the 2nd and 3rd leaf from the top were not different between nal 11 and Mianhui 727.However,width of the three functional leaves of the nal 11 was only 63％,61％ and 79％ of the width of Mianhui 727,respectively.When crossed reciprocally with three parents,the leaf width of F1 was normal.Trait segregation of leaf width occurred in all of the six F2 populations.The ratios of normal and narrow leaf plants accorded with 3:1,which suggested that nal 11 was controlled by a single recessive nuclear gene.These results lay a foundation for localization of the gene.

[目的]构建双价抗虫基因表达载体并转化水稻,为选育广谱、高抗转基因抗虫水稻提供技术参考.[方法]构建半夏凝集素基因(pta)和苏云金杆芽孢菌基因(Bf)两个抗虫基因的双价表达载体pCAMBIA 1301-Bt-pta,利用大肠杆菌DH5α和根癌农杆菌LBA4404转化水稻恢复系辐恢838,同时对转化植株进行GUS染色及PCR鉴定.[结果]以构建的双价植物表达载体pCAMBIA 1301-Bt-pta为模板,PCR扩增到pta基因的部分片段,大小为807 bp,且该载体经Hind Ⅲ酶切后,能得到15854 bp的中间载体pCAMBIA 1301-Bt片段和2046 bp的pta基因片段.对转化植株进行GUS染色,得到3株转基因阳性植株,经PCR鉴定结果显示外源基因已成功导入水稻.[结论]通过构建双价抗虫基因表达载体并转化水稻,成功获得的转基因植株可用于水稻抗虫转基因恢复系及组合的研究,以提高其抗病虫害能力.

In order to obtain good traits of rice germplasm resources, the EMS method was used to mutagenize rice Jin-hui21. Different types of stable mutants were obtained in M4 generation. Thirty-six mutants with obvious different traits were selected for SSR molecular marker analysis. Fifteen pairs of primers with stable polymorphism were screened out from 200 pairs of SSR primers covering 12 chromosomes of rice. Fifty-nine alleles were detected, effective number of alleles was 46.2574, and effective allele percentage was 78.40%. The average number of alleles per SSR locus was 3.9333 with a range of three to five. The genetic diversity index varied from 0.9951 to 1.3849 with an average of 0.9134. The polymor-phism information content for the SSR loci varied from 0.8285 to 0.9794 with an average of 0.9134. Genetic similarities a-mong the 36 mutant materials ranged from 0.515 to 1.000. The 36 mutant materials could be divided into 2 groups by u-sing the genetic similarity coefficient of 0.63.

携有包颈性状的水稻品种与花溪糯稻进行正交与反交，F1在抽穗期全部表现正常，可以正常抽穗结实，并且正交和反交2组杂交后代无明显差异，由此可以推测控制水稻包颈性状的基因为隐性细胞核基因。同时，包颈水稻突变体与花溪糯稻杂交组合F2代在抽穗期出现包颈性状植株和正常抽穗植株的分离比经卡方测验完全符合3∶1的遗传模式。由此可以推断，控制水稻包茎性状的基因为细胞核单基因隐性遗传。通过观察统计结果推测可知，控制水稻包颈性状的基因主要作用于水稻的拔节抽穗期，表达特点为水稻各茎节无法正常拔高，直接导致水稻稻穗不能正常拱出剑叶鞘，间接导致水稻穗长较短，穗粒数较少，最终导致水稻减产，但籽粒较饱满。

为了获得转基因抗虫水稻,将半夏凝集素基因pta导入水稻,并进行鉴定和筛选.在抗虫基因pta的5'端加2×35S启动子,两端分别添加HindⅢ和SpeⅠ酶切位点,并进行人工合成.将pta与植物表达载体p1301连接,经双酶切和测序鉴定.结果表明:重组表达载体p1301-pta构建成功;载体p1301-pta转化农杆菌LBA4404后用浸染法将pta基因导入水稻恢复系辐恢838中,对转化再生植株进行GUS染色以及GUS和pta基因的PCR鉴定后,共得到16株转基因阳性植株.

发掘和鉴定控制水稻株高的基因,实现对水稻株高的定向改良,具有重要的理论意义和应用价值.利用甲基磺酸乙酯(EMS)诱变水稻恢复系R30,获得了一个能稳定遗传的半矮化突变体(sd-ch).将sd-ch作母本,与“泸恢17”进行杂交,构建F2群体,进行遗传分析,并用SSR分子标记对sd-ch进行定位.结果表明,F1表现为正常亲本高度,F2群体株高出现性状分离,正常高度植株和半矮化植株数量分别为401和143,比值为2.8,经卡方检测符合3∶1,表明sd-ch受隐性单基因控制;通过SSR分子标记对sd-ch进行定位,第8染色体上SSR标记RM6028从F2群体101个隐性表型单株中检测到2个单交换株,交换率为0.99％.该基因被定位于RM6028附近,遗传距离为0.99 cM.本研究对sd-ch的初步定位,以及对其进行精心定位、克隆和应用奠定了基础.

为了研究水稻分枝突变体的遗传规律,将蜀恢527分枝突变杂合体(蜀恢527经化学诱导,M3代稳定而得)分别与正常分枝的泸恢17、R30进行正反交杂交组合,经过田间性状调查与卡方检验,发现F1代分枝突变体为隐性遗传,在F2分离群体中,分枝突变体的遗传规律符合孟德尔的常染色体单基因隐性遗传.因此,可以推断该分枝突变体是由常染色体上的隐性单基因控制.

To provide strains for biological control of rice pests,the insect-resistant gene Cry 1C from GenBank was optimized,added BamH I and Spe I restriction enzyme cutting sites at both sides,and artificially synthesized.The modified Bt gene was named Cry 1C * ,which was inserted into pTCK303 after double restriction-enzyme digestion to construct recombinant expression vector pTCK303-Cry 1C * ,and transferred into A.tumefaciens LBA4404.The PCR,double digestion,and sequencing results showed that the recombinant vector was successfully constructed(2 342 bp).The PCR result showed that the expression vector pTCK303-Cry 1C * had been transformed into A.tumefaciens LBA4404,successfully constructing a genetic engineering bacteria.

In order to study the genetic law of rice pigment genes and function of chlorophyll synthesis related genes,this study got albinos lethal mutants and chlorosis lethal mutants of R30 by mutation breeding, then used the mutants to cross and reciprocally cross with R527 and Luhui 17, analyzed the separation in F1 generation and the separation ratio in F2 generation, meanwhile, analyzed the separation ratio of heterozygous offspring. According to the result of chi-square test, the leaf color mutation of rice conforms to the genetic law of Mendel, is determined by genetic control of a single recessive gene.

This paper introduced Bt (Bacillus thuringiensis) gene and the mechanism of action of insect-resistant transgenic Bt rice, briefly summarized the research profile of Bt rice, analyzed its biological safety, and predicted its future. Therefore, it would provide references for safety evaluation and commercialization of the transgenic Bt anti-insect rice.

F 1 could be identified by DNA marker technology in genome.To provide the basis for F 2 group construction and gene location of mutants,F 1 hybridized with rice mutants were identified by SSR markers.Mutants of albefaction from R30 and of narrow leaf from ' Luhui 17 ' induced by EMS were used as parents respectively in two cross-combinations.F 1 plants from the two combinations were respectively identified by three SSR markers.The four F 1 plants(code 1-4) from mutation of albefaction from R30× ' Luhui 17 ' were identified by RM6105.The code 4 showed complementary marker type of both parents.This indicated it was real F 1.The marker types of other three were the same as its female parent.They were self-pollinated for female parent.The results of identification by RM6965 and RM5349 were consistent with that by RM6105.Three(code 9,12 and 15) from the fourteen F 1(code 5-18) from R30×Mutation of narrow leaf from ' Luhui 17 ' were identified to be real F 1.F 1 crossed with R30 and ' Luhui 17 ' could be identified respectively by the three markers of RM6965,RM5349 and RM6105 and the results were identical.F 1 identification with molecular marker could be applied in cross-breeding.

To provide materials for the research of rice functional genomics,dry seeds of ' Zhonghua11 ' and LR001 were induced with 60 Co γ-ray,then these seeds and ' Jinhui21 ' M 1 seed,induced by ethyl methane sulfonate(EMS) of 1%,were induced with 1% EMS.Through the screening to M 2,seven hundred and sixteen mutants,including the changed organ of leaf,stem,panicles and grain,etc,were identified.Among these ones,some were of great value in research,such as no-tillering,more-tillering,albino,mini-plant,thin grains,flower organs,etc.More types of mutants were obtained,with a frequencies of mutation mean of 1.79%,and all emerged in the three varieties in double induction.This indicated that the double mutation technology might provide a lot of mutants for functional genomics research.

One hundred and fifty-one rice hybrids produced in two sets of half-dialell mating crosses designed according to NCⅡ,planting respectively in Chongqing and Luzhou,Sichuan Province of China in 2005 were used to predict the F1 performances of seven yield traits through genetic effects of marker loci of AFLP and SSR.Positive loci(PL)and effect-increasing loci(IL)were screened respectively by yield traits of two sets of crosses and their additive or/and dominant effects were used to founded the prediction model for these yield traits.Predictions of yield traits of crosses derived from different parents(prediction between sets)and fixed parents were conducted by the model founded.The results were as follows:(1)Most predictions between sets were not ideal and changeable.The correlations between predictive values and actual ones ranged from-0.55 to 0.45.(2)The prediction effects of crosses with by PL were similar to those by IL in Set 2,and all better than that of between sets.Predictions of crosses with fixed restorers were more valuable than those of crosses with fixed sterile lines.(3)Most predictions of crosses with fixed parents were relatively better.The predictions by IL of seed setting rate of crosses with fixed sterile lines,panicle number of those of fixed restorers,and those by PL of spikelets per panicle and grain weight per panicle of crosses with fixed restorers were above 0.6.Because the number of PL and IL in prediction model were much less,the prediction models was conveniently applied and particularly,and it was important for the institute by which the models were founded in parent-selection in breeding.

The positive, effect-increasing and effect-decreasing loci (PL, IL, DL), which were screened from the parental AFLP polymorphic loci (TL) by the yield traits of two sets of half-dialell mating crosses designed used 13 cytoplasmic male sterile lines and 19 restorers of rice, respectively, together with TL, were utilized to estimate parental genetic distances (GDs), and the models were founded to predict the traits of hybrids derived from different parents. The results were as follows: 1) PL occupied a few number of the TL, and its correlations between the predictive and authentic values of yield traits (termed as briefly predictive coefficient as following) were too low, as same as TL. 2) But the predictions by IL, DL or IL/DL (founding duality models) were increased significantly, e.g. the predictive coefficients were all above 0.6 such as IL for grain setting percentage, IL, DL or IL/DL for grain weight per plant, therefore, these models could be used as prophecy of rice breeding. 3) In addition, the predictions based on IL, DL or IL/DL from intersection of two sets of crosses were higher enough, four of which above 0.5, and was more suitable in molecular breeding due to less predictive loci required.