Attaining inter-subgeneric hybrids in fragrant azalea breeding and the inheritance of organelle DNA

Inter-subgeneric hybrids were successfully obtained in reciprocal cross combinations between evergreen azaleas (Rhododendron nakaharae and its hybrids) and fragrant deciduous azaleas (R. arborescens and R. viscosum) for the purpose of fragrant evergreen azalea breeding. Nuclear and organelle DNA of these hybrids was investigated using PCR-RFLP markers. Viable hybrid seedlings have nuclear ribosomal DNA (nrDNA) inherited biparentally, mitochondrial DNA (mtDNA) from the seed parent, and chloroplast DNA (cpDNA) from the deciduous azalea, regardless of cross combination. These results suggest that the chloroplast genome from deciduous azaleas and the nuclear genome from evergreen azaleas are compatible in viable hybrid progenies.     

Appearance of albino seedlings and ptDNA inheritance in interspecific hybrids of azalea

PCR-RFLP analysis was conducted to clarify the relationship between the leaf color of progenies and their ptDNA inheritance in interspecific three-way crosses, (Rhododendron kiusianum × R. eriocarpum) × R. japonicum f. flavum. All albino progenies contained maternal ptDNA, whereas green and pale-green progenies contained paternal ptDNA. Sectorial chimeric progenies, of which the leaf and shoot color was turned from green to albino during the culture, contained both maternal and paternal ptDNA in green segments and maternal ptDNA in albino segments. These results suggest that albino progenies are caused by the incompatibility between plastome from F₁ hybrids of R. kiusianum × R. eriocarpum and nuclear genome from R. japonicum f. flavum. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effectiveness of three-way crossing for the breeding of yellow-flowered evergreen azalea

For the purpose of obtaining yellow-flowered evergreen azaleas, single and three-way interspecific crosses between evergreen azaleas and Rhododendron japonicum f. flavum were compared. In single cross, although many seeds were obtained from R. eriocarpum × R. japonicum f. flavum, the seedlings from immature ovule were albino or pale-green plants. They were withered away at acclimation. In R. kiusianum × R. japonicum f. flavum, green seedlings were observed, but number of ovules contained in one capsule was very small. In three-way cross, (R. kiusianum × R. eriocarpum) × R. japonicum f. flavum, many viable seeds were contained in one capsule, and about 15% of the seedlings were green leaf colored. These results suggest that three-way cross is one of the useful breeding methods for creating the new yellow-flowered evergreen azalea. Isozyme analysis revealed that most of seedlings were zygotic origin without distinction of their leaf colors, but some green seedlings indicated the same genotypes as the seed parents. Therefore, it is important to confirm hybridity of the seedlings by isozyme technique. This revised version was published online in August 2006 with corrections to the Cover Date.     

Development of multiplex PCR markers to distinguish evergreen and deciduous azaleas

We successfully developed new PCR‐based markers for analysing the DNA inheritance patterns of vigorous inter‐subgeneric hybrids of azalea. The hybrids were between evergreen azaleas (Rhododendron nakaharae and its hybrids) and fragrant deciduous azaleas (Rhododendron arborescens and Rhododendron viscosum). We determined the sequence of the nuclear internal transcribed spacer (ITS) region in evergreen and deciduous azaleas, and found a deletion of five base pairs (bp) in evergreen azaleas compared with deciduous azaleas. Sequences for the chloroplast matK‐trnK region were obtained from GenBank. Primers for the multiplex PCRs were designed using these sequence data. In the multiplex PCR of the ITS region, evergreen azaleas showed a specific band of 140 bp and deciduous azaleas showed a specific band of 800 bp. In the multiplex PCR of the matK‐trnK region, evergreen azaleas showed a specific band of 560 bp and deciduous azaleas showed a specific band of 300 bp. These results indicate that multiplex PCR methods are useful to distinguish evergreen and deciduous azaleas easily and rapidly.     

Development of advanced fragrant rice lines from MR269 × Basmati 370 through marker-assisted backcrossing

Fragrance in rice is an appealing attribute to consumers. The increasing demand for fragrant rice highlights the need to develop fragrant rice variety that suit the preference of local consumers in addition to reduce fragrant rice imports. Marker-assisted backcrossing (MABC) was employed to develop advanced fragrant rice lines from the cross between MR269 and Basmati 370. MR269 is a Malaysian high-yielding rice variety but non-fragrant and was used as recurrent parent whereas Basmati 370 is a well-known fragrant traditional rice variety and was used as donor parent for the fragrance gene. Two generations of backcrosses and a generation of selfing were conducted to introgress the fragrance gene and restore the recurrent parent genome in the backcross progenies. As a result, 14 advanced fragrant rice lines were developed. These advanced fragrant rice lines carried homozygous alleles for the fragrance gene, similar to Basmati 370. The average recovery of recurrent parent genome was 88.4%. Besides being fragrant, the advanced fragrant rice lines also had most of the morphological and agronomical traits similar to MR269. Grain quality of the advanced fragrant rice lines in terms of gelatinization temperature, amylose content and gel consistency are also similar to both parents. Besides, the advanced fragrant rice lines had 2-acetyl-1-pyrroline content similar to Basmati 370. MABC approach applied in this study has successfully introgressed the fragrance gene and accelerated the recovery of recurrent parent genome in advanced fragrant rice lines, therefore these lines can be delivered to the farmers and consumers for use in due time.     

Linkage between genes for hairy first leaf and chlorophyll deficiency in Brassica oleracea

The fourth linkage group of B. oleracea L. has two genes: Hr-1, (hairy first leaf), a dominant seedling marker from “Dwarf Green” curly kale, and pg-2, (pale green seedling), a recessive chlorophyll mutant from green sprouting broccoli. Recombination between Hr-1 and pg-2 ranged from 7.4 to 20.1% in the six progenies studied, with a mean of 13.15±0.68%. Hr-1 segregated independently of the three other linkage groups (two genes of each were tested) and of two unlocated genes for male sterility.     

柠檬合子胚自交实生苗的鉴定及其对柑橘溃疡病的抗性初步评价

枸橼C-05是抗溃疡病的柑橘种质资源,柠檬是枸橼的后代,其自交后代可能继承枸橼的抗性基因。为分析柠檬自交群体对柑橘溃疡病的抗感性规律,本研究建立了433株的‘尤力克’柠檬自交后代群体,并利用SSR分子标记对合子胚实生苗进行了鉴定,观察合子胚实生苗和珠心胚实生苗叶片形态学的差异,对所有自交后代离体注射接种柑橘溃疡病菌,初步评价了实生后代对柑橘溃疡病的抗病性差异。结果表明‘尤力克’柠檬存在多胚性,从300对SSR引物中筛选到8对可用于‘尤力克’柠檬合子胚鉴定的引物,共鉴定出合子胚实生苗57株,确定珠心胚实生苗4株。叶形指数、叶尖指数、翼叶长、翼叶宽和叶柄长均不能作为区分‘尤力克’柠檬合子胚实生苗与珠心胚实生苗的形态学标记,但合子胚实生苗的翼叶长度变异幅度大于珠心胚。57株合子胚实生苗离体接种柑橘溃疡病菌后,5株表现为抗病,52株表现为感病;4株珠心胚实生苗均表现为感病;在372株未鉴定清楚的自交后代中,8株表现为抗病,364株表现为感病。本研究表明利用SSR分子标记还不能完全鉴定出‘尤力克’柠檬合子胚植株,可能需要更多的引物或更灵敏的分子标记才能把全部实生后代鉴定出来。利用合子胚自交后代接种鉴定,更有利于分析‘尤力克’柠檬对柑橘溃疡病的抗感性遗传规律,挖掘抗性基因,为柠檬的抗病育种提供种质资源。     

Interspecific hybridisation between Hibiscus syriacus, Hibiscus sinosyriacus and Hibiscus paramutabilis

Interspecific hybrids from crosses between H. syriacus × H. paramutabilis and H. syriacus × H. sinosyriacus were obtained. In both cases unilateral incongruity was observed and reciprocal crosses yielded no fruits. In vitro embryo rescue, 11 weeks after pollination, increased the germination capacity of H. syriacus × H. sinosyriacus embryos, while this was not the case for H. syriacus × H. paramutabilis embryos. However, a lot of the generated H. syriacus × H. sinosyriacus seedlings were lost due to variegated and total albinism. In the progeny of H. syriacus ‘Oiseau Bleu’ × H. paramutabilis about 95% of the seedlings had an intermediate flower and leaf morphology compared to both parent plants. Leaves on the adult F1 hybrids showed a polymorphism. In total four different leaf types could be observed on the same plant. This leaf polymorphism also was seen in the progeny of H. syriacus ‘Melwhite’ × H. sinosyriacus ‘Lilac Queen’. In this progeny about 50% of the seedlings had an intermediate flower and leaf morphology compared with the parent plants. The hybrid nature of the seedlings of both progenies was also confirmed by AFLP analysis. Despite the low germination vigour of the pollen of the hybrids, a small F2 generation was obtained from H. syriacus ‘Oiseau Bleu’ × H. paramutabilis.     

大豆细胞质遗传芽黄突变体的发现

本研究新发现一个大豆芽黄突变体NJ89-3。该突变体典型表现为，从第一对真叶开始，新生叶表现明显黄化，但随叶片充分伸展定形，逐渐变为正常绿色。植株最后一张叶片充分伸展定形后，芽黄植株与正常绿色植株并无明显区别。研究表明NJ89-3的芽黄性状由细胞质基因突变引起，属细胞质遗传，建议将此细胞质基因命名为cyt-Yv。     

水稻叶缘白化突变体mal的遗传分析与基因定位

植物叶色变化对叶绿体发育和叶绿素生物合成等光合系统结构和调控机制的研究有着重要的理论意义。水稻叶缘白化突变体mal (marginal albino leaf),来源于恢复系缙恢10号(Oryza sativa L.ssp. indica)的EMS诱变群体,经过多代自交,其突变性状遗传稳定。与野生型相比,mal突变体整个生育期叶片边缘白化且叶片变窄,抽穗期倒三叶叶片、倒二叶叶边缘以及倒三叶叶边缘的叶绿素含量极显著降低。透射电镜观察发现,mal突变体叶片绿色部位细胞与叶绿体发育完全,白化部分叶肉细胞大部分中空,无明显完整的细胞器,叶绿体内部完全降解。遗传分析表明该突变体受隐性核基因控制,MAL被定位在第8染色体上SSR标记M22和InDel标记ID27之间,物理距离为171 kb。本研究将为MAL基因的图位克隆及功能研究奠定基础。     

EMS诱变甜高粱突变体筛选与鉴定

为了创制优良甜高粱种质资源,改良高配合力父本性状,为甜高粱遗传育种奠定基础,本试验利用24 h+0.25%甲基磺酸乙酯(EMS)水溶液诱变甜高粱甜C-1,从M2代中筛选鉴定优良突变体用于下一步遗传育种。研究表明叶色突变主要发生在M1代,且大部分不能遗传,M1代发现的突变,M2代中依然能够大量发现同类突变。在M2代田间鉴定中发现的突变体类型主要有叶色白化和黄化突变、穗型突变为纺锤型、颖壳包被度变异为1/2和3/4包被、颖壳颜色突变为黑褐色、籽粒增大、籽粒突变为白色、芒性消失、全生育期缩短。利用EMS诱变可以改良现有优良父本性状,加速杂交种的选育,且发现大量的突变体对甜高粱基因功能的挖掘和遗传育种有积极的意义。     

Segregation of Isozyme Marker Loci Amongst Androgenetic Plants of Lolium perenne L.

The segregation of isozyme markers has been examined in the green and albino androgentic progenies of two plants of Lolium perenne. Overall the pooled data for the loci PG1/2 and GOT/2 does not deviate from expectation, but there is considerable heterogeneity between families and between green and albino plants. Specific alleles predominate in the different progeny sets. The results may be accounted for by possible linked loci affecting anther culture response or of linked lethal genes.     

绿豆几个表型性状的遗传特性分析

种皮色泽和花青甙显色是绿豆(Vigna radiata)的重要性状,可在纯度鉴定、辅助选择育种等研究中发挥作用。本研究以绿豆核心种质种皮色泽、花青甙显色性状分析为基础,配制杂交组合,根据不同世代的表型调查,探讨绿豆种皮颜色、光泽和花青甙显色等性状的遗传特征。结果表明,绿种皮对黄种皮为单基因控制,绿色为显性;黑种皮对绿种皮为单基因控制,黑色为显性。绿豆种皮光泽不符合简单性状的遗传规律,且不同组合间后代的表型分布存在差异。绿豆种皮色与光泽间并无连锁关系。植株各部位的花青甙显色对不显色为显性,均符合简单单基因的分离规律。进一步分析表明,不同部位花青甙显色的相关性不一致,其中控制幼茎色基因与控制复叶基部花青甙显色基因紧密连锁在一起,或为一个基因,而控制龙骨瓣花青甙显色的基因与其他相关基因的遗传连锁程度最低。本文对绿豆种皮相关性状及植株花青甙显色性状的遗传分析将为这些性状的育种利用及深入研究奠定基础。     

Development of elite rice restorer lines in the genetic background of R022 possessing tolerance to brown planthopper, stem borer, leaf folder and herbicide through marker-assisted breeding

Rice leaf folder, stem borer and brown planthopper (BPH) are the most devastating rice insect pests. Developing and planting insect-resistant rice varieties is the most economical and effective measure for controlling these pests. BPH can be controlled with native BPH-resistance genes in rice, while at present rice leaf folder and stem borer can only be controlled through planting transgenic Bt rice. In this study, the breeding of a new restorer line KR022 possessing stacked BPH-resistance genes Bph14 and Bph15, Bt gene cry1C and glufosinate-resistance gene bar, is reported for the first time. A rice restorer line R022 with BPH-resistance genes Bph14 and Bph15 was used as a recurrent parent to cross with the transgenic rice T1C-19 of cry1C and bar genes during the breeding process. The restorer line KR022 was developed from the backcross populations of R022 and T1C-19 through molecular marker-assisted selection and glufosinate-resistance selection. The cry1C and bar genes were found to integrate on chromosome 11 of KR022, and the genome recovery of KR022 was up to 95.8 % of the R022 genome. The quantification of Cry1C protein expression showed that it was expressed at different levels in the leaf, stem, panicle, endosperm, and root of KR022 and its hybrid rice. The insect-resistance evaluation indicated that KR022 and its hybrid rice had good resistance to rice leaf folder and stem borer, both in laboratory settings and in the field. Furthermore, they exhibited increased resistance to BPH at both the seedling and mature stage. The field trial showed there was no significant difference in key agronomic traits between KR022 and its recurrent parent R022, and four hybrids from KR022 yield much higher than the control II-You 838. Moreover, KR022 and its hybrid rice were found to have resistance to the herbicide glufosinate. These results demonstrate that KR022 is effective as a rice restorer line for the breeding of “green super rice”, possessing multiple tolerances to rice BPH, stem borer, leaf folder and glufosinate.     

小麦叶绿素缺失突变体Mt135的叶绿体基因差异表达分析

小麦叶绿素缺失突变体Mt135自交后代稳定表现绿株、条纹株和白化株3种类型, 其中条纹株白色组织和白化株的叶绿体数目和结构发生突变, 完全失去光合能力。为研究该突变体叶绿体基因表达与光合作用的关系, 采用实时荧光定量PCR技术, 分析了白化株和条纹株的叶绿体基因表达。在白化株中共检测到40个差异表达基因, 涉及4类功能(编码光反应相关蛋白、编码叶绿体内能量代谢相关酶、核糖体合成和tRNA合成), 包括18个上调表达和22个下调表达基因;在条纹株中共检测到13个上调表达基因, 其表达变化趋势与在白化株中一致。白化株的差异表达基因中, 编码光系统II、I结构蛋白的psb、psa及ycf等基因家族的基因表达量显著下调;多个编码核糖体蛋白大、小亚基的基因表达量改变, 尤其是核糖体蛋白小亚基编码基因rps14和23S rRNA的编码基因23S rDNA表达量显著下调。推测Mt135突变性状与参与光反应相关蛋白的编码基因、叶绿体内能量代谢相关酶的编码基因、核糖体合成相关基因以及tRNA合成相关基因表达量的改变密切相关。     

Inheritance of tree and fruit characters in progenies from crosses of sweet cherry (Prunus avium L.) cultivars

Summary Approximately 1000 seedlings from 20 combinations crossed in 1979, 1980 and 1981 (Theiler-Hedtrich 1985a) were tested for several characters: fruit set (yield), fruit size, fruit colour, formation of abscission layer and bleeding after fruit removal from fruit stalks, bacterial canker resistance, flowering and harvesting time. From progeny of crosses with Stella as pollinator, 56% (Vittoria × Stella) and 46% (Schüttler × Stella) of the seedlings were self-compatible, of which 14 were high yielding with good fruit size and quality. From the data recorded it can be concluded:fruit set is a recessive character; only 5 to 20% of very good yielding seedlings were obtained in different progeny, even if the parental plants were both very good croppers.Fruit juice and skin colour was in most progenies ‘black’ even if they were from combinations with ‘white’ varieties, e.g., Merton Glory or Schüttler. Only from the combination Schiittler (‘white’) × Stella (‘black’), 50% of the seedlings were ‘white’; Stella therefore is heterozygous for the character of fruit juice and skin colour.Fruit size is evenly distributed in progeny with respect to the fruit size of their parent plants.Abscission layer formation and non-bleeding is a genetically complex character. In combinations where both parent plants formed fruits with complete abscission layers and which were not bleeding after fruit removal from the stalk, this character was inherited only to 50% (Vittoria × Schüttler) or 85% (Vittoria × Frühe von der Weid) in the progeny. For the genetical control of this character further studies are necessary.Bacterial canker susceptibility was evenly distributed in seedlings from all combinations even if the highly resistant cv. Vittoria was used as one parent plant, thereby not confirming the expected results of a higher proportion of resistant seedlings from combinations with Vittoria.Flowering and harvest time of the seedlings from different combinations was within the range of the parent plants. Only in the combination of Vittoria × Stella (mid-to late-ripening season) one seedling out of 99 was found to form ripe fruits two weeks earlier than the parental plants. From the seedlings tested 40 have been chosen for further evaluation or genetical studies.     

Genetic analysis for haploid-regenerations responses of the hexaploid-wheat anther cultures

Genetic variability in response to anther culture was investigated in 49 winter hexaploid wheats, comprising 33 pure lines (F₁₀) derived from a composite cross programme and their 16 parental genotypes. All genotypes were grown in a randomized block design with three replications in a controlled greenhouse. The number of embryoids and total plant regeneration per 100 anthers, as well as the numbers of green and albino plants regenerated per 100 embryoids, were measured. Significant genetic variability was observed among the 49 genotypes for all the traits studied. All traits showed high heritability. Among the genotypes compared, DC²30N and 1BPT-40 gave the best results for the production of embryoids and IBPT-78 had the highest value for the production of green plants. The genotype IBPT-34 developed a large number of albino plants, and it should be useful as a parent in studies to determine the genetic control of albino plants in wheat.     

水稻耐低磷种质的苗期筛选与鉴定

从以成恢448为轮回亲本，M401为供体亲本构建的回交后代BC₁F₆筛选出的耐低磷材料（GP9）和低磷敏感材料（GP14）及其亲本用于盆栽试验，设置P2（2.31 mg/kg）、P30（30 mg/kg）和P80（80 mg/kg）3个磷处理水平，以P80处理为对照计算性状相对值，研究不同磷处理浓度对供试材料6个生物学性状的影响。结果表明，P2处理较P30处理更能反映不同材料性状相对值间的差异，宜用于苗期耐低磷筛选。P2处理时，GP9与GP14及其亲本播种后52 d、59 d和66 d的相对分蘖数差异均达极显著水平，建议采用相对分蘖数作为水稻苗期耐低磷筛选的主要鉴别指标。同时，运用相对叶面积、相对绿叶数和相对叶龄进行综合评定。     

澳洲坚果人工杂交子代苗木生长量变异分析

为了认识澳洲坚果植株生长量性状的传递遗传规律,以澳洲坚果10个无性系品种为材料,正反交设计,共90个杂交组合,获得90个全同胞家系苗木。并对种子横径、苗木株高、地径及冠幅进行变异分析。结果表明,10个母本间种子横径达显著差异(P<0.05),同一母本所对应9个父本的组合间种子横径达显著(P<0.05)或极显著(P<0.01)差异,说明种子的大小不仅仅决定于母本,而且也受父本的影响。对苗木株高、地径和冠幅的分析表明,全同胞家系间差异显著(P<0.05),母本半同胞家系间差异显著(P<0.05),而父本半同胞家系间差异不显著。苗木生长量受父母本的基因共同决定,但母本影响的变异分量相对较大,父本影响的变异分量相对较小。