毛新杨×毛白杨叶片表型和春季萌芽时间QTL分析

以毛新杨无性系TB0 1×毛白杨无性系LM5 0的回交子代 12 0株个体为作图群体 ,对控制叶片表型性状如叶长、叶宽、叶面积和叶柄长以及春季萌芽时间共 5个性状的数量性状位点 (quantitativetraitloci,QTLs)进行分析。运用AFLP标记技术结合拟测交作图策略构建了含有 393个AFLP标记的毛白杨及毛新杨的遗传连锁框架图。毛白杨遗传图上共含有 2 4 7个AFLP标记位点 ,连锁位点覆盖毛白杨基因组总长约 32 6 5 1cM ,而毛新杨遗传连锁图上共含有 14 6个标记位点 ,连锁位点覆盖毛新杨基因组总长约 1992cM ,这些连锁图的每一连锁群上含有的标记数为 4～ 30个。在此基础上 ,利用区间作图软件共检测到控制叶片表型性状的QTLs14个 ,位于 9个连锁群上 ;而对于春季萌芽时间共检测到 3个QTLs,分别位于 3个不同的连锁群上。在检测到的 17个QTLs中 ,每一QTL可解释表型变异的 7 6 %～ 15 8%。此外 ,发现控制叶长、叶宽和叶面积等相关性状的QTLs位于相同的基因组区域 ,这些QTLs主要位于毛白杨遗传连锁图的TLG2和TLG11以及毛新杨连锁图的TBLG1连锁群上。据控制叶片表型和春季萌芽时间的QTLs所处的基因组区域 ,可推测叶片表型和春季萌芽时间这两类性状是由各自相应的基因控制。     

Duplication of locus coding of malate dehydrogenase in Populus tomentosa Carr.

Horizontal starch-gel electrophoresis was used to study crude enzyme extraction from young leaves of 234 clones of Populus tomentosa Carr. selected from nine provenances in North China. Ten enzyme systems were resolved. One hundred and fifty-six clones showing unusual allozyme band patterns at locus Mdh-1 were found. Three allozyme bands at locus Mdh-1 were 9:6:1 in concentration. Further studies on the electrophoretic patterns of ground mixed pollen extraction of 30 male clones selected at random from the 156 clones were conducted and it was found that allozyme bands at locus Mdh-1 were composed of two dark-stained bands and a weak band. Only one group of the malate dehydrogenase (MDH) zymogram composed of two bands was obtained from the electrophoretic segregation of pollen leachate of the same clones. A comparison of the electrophoretic patterns one another suggested that the locus Mdh-1 coding malate dehydrogenase in diploid species of P. tomentosa was duplicated. The duplicate gene locus possessed three same alleles and was located in mitochondria. The locus duplication of alleles coding malate dehydrogenase in P. tomentosa was discovered and reported for the first time. [Supported by the “Tenth Five-year Plan” National Key Project in Science and Technology (Grant No. 2002BA515B0303) and the National “863” Project (Grant No. 2002AA241071)]     

TCF bleaching of <Emphasis Type="Italic">Populus tomentosa</Emphasis> kraft pulps with H<Subscript>2</Subscript>O<Subscript>2</Subscript> pretreatment under acid condition

The process of peroxide bleaching of Populus tomentosa kraft pulp with H₂O₂ pretreatment under acid condition was studied. The variations of pulp characteristics such as brightness, Kappa number and viscosity during the peroxide bleaching process were discussed and the concept of the A/B value was presented for the first time. The results show that acid pretreatment with H₂O₂ is essential to improve pulp brightness and avoid viscosity loss. The conclusion is reached that the A/B value has great influence on pulp properties when the total amount of peroxide used in the bleaching process is 2%–5%. Pulp with a high viscosity and brightness can be obtained when A/B is about 0.15. [Supported by the National “863” Project (Grant No. 2002AA241071)]     

Variation of leaf characteristics in Populus tomentosa Carr.

An investigation was conducted to determine the extent of variations among nine provenances of Populus tomentosa Carr. in terms of leaf characteristics. A total of 263 accessions were studied under field conditions in the National Gene Bank of P. tomentosa in 2003. All of the accessions were characterized by 17 indices from 1 to 2-dimension constructions. Variance analysis of all characteristics showed that there were significant differences among the nine provenances and among individuals within each provenance. This study reveals that the evaluated germplasm appears to have a wide genetic base and high potential for further genetic improvements and it also indicates that abundant gene resources of P. tomentosa have been collected and preserved in the National Gene Bank. [Supported by the “Tenth Five-year Plan” National Key Project in Science and Technology (Grant No. 2002BA515B0303) and the National “863” Project (Grant No. 2002AA241071)]     

Genetic variation ofAlnus trabeculosa populations in Japan

Alnus trabeculosa Hand.-Mazz. is classed as “near threatened,” and efforts should be made to conserve the species, but effective action would require understanding of its genetic variation and structure. Therefore, the genetic variation at 12 isozyme loci was examined in seven populations of the species in Japan. The expected heterozygosity within species (H _es=0.222) and the expected heterozygosity within populations (H _ep=0.199) were found to be slightly higher than in most other woody species. The coefficient of genetic differentiation (G _ST=0.146) was also high. The northernmost population had the lowest values for almost all measures of genetic variation, and populations in southern Japan tended to show greater within-population variation than those in northern Japan. The level and distribution of genetic variation in this species might be related to its life history and ecological traits, and distribution history.     

Adaptive genetic diversity and population structure of the “algarrobo” [<Emphasis Type="Italic">Prosopis chilensis</Emphasis> (Molina) Stuntz] analysed by RAPD and isozyme markers

The “algarrobo” [Prosopis chilensis (Molina) Stuntz] is a tree species that represents an important natural resource in arid and semi-arid regions of Argentina. In this paper, we analysed and compared the variability of 46 RAPD (Random Amplified Polymorphic DNA) loci with previous estimates obtained from 12 isozyme markers in nine Argentinean populations of P. chilensis representative of the whole range of this species in Argentina. We evaluated the population structure and the existence of genetic variants associated with environmental variables. Expected heterozygosity (H _e) estimated from RAPD varied significantly among populations and regions. Hierarchical analysis of genetic variability (AMOVA) showed that most (88.1%) of the total diversity occurs within populations, the component among populations within regions (9.3%) was intermediate, while the between-region component was the lowest (2.6%). All three variance components were highly significant. The MDS plot from pair-wise Φ _ST matrix was consistent with the highly significant among-region differentiation indicated by the AMOVA. All 12 variable isozyme loci and 26 out of 46 RAPD loci showed significant or highly significant association with at least one geographic/climatic variable according to the stepwise multiple regression analysis. These results imply that the genetic differentiation among populations is better explained by environmental or biogeographical grounds than by geographical distances, suggesting gametic disequilibrium with loci responsible for the adaptation to particular environmental conditions. The information from RAPD markers would provide a relevant criterion to preserve genetic diversity in programmes for conservation and rationale use of this species.     

Die Vererbung der Resistenz von Pappeln gegenüberGypsonoma aceriana Dup. undPhyllocnistis suffusella Z.

Zusammenfassung Die Resistenz von Pappeln, die in vorliegenden Untersuchungen verwendet wurden, gegenüberGypsonoma aceriana undPhyllocnistis suffusella scheint unter strenger genetischer Verankerung zu stehen. Die Daten zeigen, da? die Eigenschaften der Resistenz sich recessiv in heterocygoter Kombination mit den sensitiven verhalten. Klon „ I-455” scheint resistenter als Klon „I-214” gegen Befall vonGypsonoma aceriana zu sein.

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Summary The resistance of Poplars used in this experiment toGypsonoma aceriana andPhyllocnistis suffusella appeared to be under strong genetic control. It was presented data showing that the character for resistance behaves as recessive in heterojugous combination with the sensitive one. Clone “I-455” appeared to be more resistant than clone “I-214” toGypsonoma aceriana attack.

     

First results on synchrony between seasonal pattern of pheromone trap captures of cotton bollworm, <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> and appearance of freshly emerged larvae on developing cobs of corn hybrids

Synchrony between development of five corn hybrid varieties of various seasonal growing rates (FAO numbers), seasonal flight pattern of male cotton bollworm, Helicoverpa armigera Hb. (Lepidoptera: Noctuidae), as monitored by pheromone traps, and the subsequent appearance of newly hatched larvae on developing cobs were studied at Mezőhegyes, South-Hungary, in 2003 and 2004. The phenological stages of corn hybrids were evaluated using the Iowa State University Scale (R₁–R₅), the flight of male moths was monitored by large capacity, funnel type of pheromone traps and the appearance of freshly hatched larvae on developing cobs were counted by visual inspection. The synchrony between the flight peak of male moths and the peak apperance of L₁ larvae on cobs was investigated by cross-correlation. In 2003 (average daily temperatures 20.7°C, average daily relative humidity 59.9% for the period of 4–18 July) “DK 391”, “DK 443” and “Maraton” hybrid varieties already reached silking stage (R₁) by the time when pheromone traps indicated a peak, at 7 July (peak capture at 3-day intervals 755.5 males/traps). The number of L₁ larvae peaked on developing cobs of these varieties also at 7 July (7.0, 4.0 and 3.8 larvae/50 cobs, respectively). The synchrony between the flight peak of male moths and the peak appearance of L₁ larvae on cobs was proven (LAG = 0). A rather similar trend was observed in “Vilma” hybrid variety: it reached R₁ stage at 7 July, and L₁ larvae appeared only a few days later (11 July, 3.8 larvae/50 cobs) (LAG = −1). However, “Maxima” hybrid variety reached R₁ stage 1 week later (14 July) than the time of peak captures. Here L₁ larvae peaked as late as at 18 July (0.8 larvae/50 cobs), i.e., only after the cob had reached R₁ stage (LAG = −3). A reverse order of dates of R₁ stage and peak capture was observed in “Maxima” in 2004 (average daily temperatures; 22.5°C, average daily relative humidities 72.6% for the period of 15 July–6 August): it reached R₁ stage at 19 July, while peak trap captures were recorded at 6 August (peak capture at 3-day intervals 20.5 males/traps). L₁ larvae were found in the highest numbers on 2 August (1.5 larvae/50 cobs), practically in synchrony with peak caputres (LAG = 0). “Káma” reached R₁ stage in 16 July, and L₁ larvae peaked at 2 August (1.3 larvae/50 cobs) (LAG = 0). On the rest of the corn varieties larvae were found only in too numbers for performing statistical analysis. We conclude that in order to predict the appearance of L₁ larvae, the phenological stage of the corn variety and the seasonal flight pattern of moths, as measured by large capacity pheromone traps, should be considered in combination. If the corn variety already reached R₁ stage, L₁ larvae appear on cobs as early as the time of peak flight of moths. However, no young larvae appear on cobs despite of high trap captures, until the corn reaches the R₁ stage. These findings are discussed in the view of specifying optimal timing of a pesticide application.     

Marker trait association for autumn cold acclimation and growth rhythm in pinus sylvestris

The main objective of this study was to identify quantitative trait loci (QTLs) involved in the genetic control of growth rhythm and autumn cold acclimation in Pinus sylvestris L. Eighteen full‐sib families of P. sylvestris with parental origins between latitudes 62 and 67° N in Sweden were assessed for autumn cold acclimation (frost injury measured from artificial freeze testing) and growth rhythm during the first two growing seasons. One family north × south with 108 individuals was used for the construction of one linkage map for the male and one for the female parent using 286 markers from random amplification of polymorphic DNA. Marker trait associations (QTLs) for shoot elongation, growth cessation and cold acclimation were found on both maps. Some QTLs were verified in two additional north × south families. Proportions of the additive genetic variance explained by the markers (R ²/_G) indicated that genetic factors of large effect were involved in all investigated traits. Single or multiple markers accounted for between 16 and 47% of the additive variance in annual shoot elongation, between 42 and 79% of the additive variance in growth termination, and between 7 and 31% of the additive variation in autumn cold acclimation. The study confirms that there are major QTL regions on different linkage groups controlling a large part of the variation for growth rhythm and autumn cold acclimation in P. sylvestris.     

RAPD markers related to sex locus in Populus tomentosa

By using the methods of random amplified polymorphic DNA (RAPD) and bulked segregate analysis (BSA), we identified markers that are linked to the sex determination in the dioecious Populus tomentosa. Male and female bulks were created through rough mixing equal amounts of its five individual DNA. A total of 88 primers were screened. Twelve primers produced clear patterns with at least one band that appeared to be polymorphic between the two bulks. Subsequently, five male and female individuals were analyzed with those 12 primers, and only S60 (ACCCGGTCAC) could generate a common 1800 bp DNA fragment in all five male individuals and male pool but not in any female individuals. It can be concluded that the gender of P. tomentosa is most likely connected to the S60-1800 bp DNA fragment and RAPD markers. S60, therefore, can be used for selecting the gender of P. tomentosa. __________ Translated from Journal of Central South University of Forestry & Technology, 2008, 28(3): 80–83 [译自: 中南林业科技大学学报]     

Cross breeding of Populus and its hybrids for cold resistance

Populus tomentosa was crossed with P. tremuloidis, P. grandidentata, P. alba × P. grandidentata and P. alba × Ulmus pumila in order to maintain its rapid growth and high wood quality and improve its resistance to cold. Two methods were used to increase the germination rate from 1.5% to 41.1% and the remaining rate from 1.7% to 44.2%. Forty crossing combinations were conducted and 2 744 hybrid seedlings were obtained. MX₄ × P. grandidentata (G-1-58), MX₃ × P. tremuloidis (T-44-60), MX₂ × P. tremuloidis (1-13-87-37) and MX₂ × (P. alba × P. grandidentata) were regarded as superior combinations after analysis and selection. Thirty seedlings of these combinations and 11 triploid seedlings identified by counting their chromosomes were selected as super plants. [Supported by the Doctoral Foundation of Ministry of Education (Grant No. 200220022004) and the National “863” Project (Grant No. 2002AA241071)]     

Webbing clothes moth,Tineola bisselliella (Hum.), causing serious feeding damage toLactuca sativa and other plant seeds

The first record of a “field” infestation of stored lettuce seeds (Lactuca sativa) byTineola bisselliella (TB) is reported. The pest is able to consume almost the whole kernel ofL. sativa including the shell. The ability od TB to damage selected stored cereal, pulse and oil seeds under laboratory condition was evaluated. In maize and wheat, germ area was damaged preferably and the germination decreased considerably. TB was unable to infest and damage sound seeds of rape and beans ofPhaseolus vulgaris and soya. These findings enabled the development of a vegetarian diet for laboratory rearing of TB. The pest status and hazards resulting from cross-infestation is discassed.     

Patterns of genetic diversity that result from bottlenecks in Scots Pine and the implications for local genetic conservation and management practices in Bulgaria

As one of the principal coniferous species of commercial importance in Bulgaria, Scots Pine has been subjected to strict management, which has posed a challenge for the conservation and management of its gene pool. In this study, we measured genetic diversity and population structure using 6 microsatellite loci in 12 populations from Bulgarian mountain ranges. Our analysis showed good intrapopulation divergence (18.22%; AMOVA) and genetic evidence of bottlenecks. “NJ-IAM” and “non-admixing” clusters divided the populations into 3 large groups. Our microsatellite analysis was consistent with the documented “mountain effect” influence on genetic diversity. Two methods—the Coalescent Bayesian model and M-stat analysis—implicated recent bottlenecks for all populations, suggesting that the genetic structure of Bulgarian Scots Pine was shaped over a long period (one thousand generations; i.e. 20,000 years) by the interaction of genetic and autoecological processes. Our results suggest that conservation and management practices should strive to maintain this genetic differentiation, specifically by emphasizing reforestation efforts with stocks from local provenances to avoid nonlocal introductions.     

Molecular cloning of a cellulose synthase gene PtoCesA1 from Populus tomentosa and its genetic transformation in tobacco

A 3 125 bp cellulose synthase gene, PtoCesA1, which has a 98% identity to PtrCesA1 from Populus tremuloides, was cloned from cDNA prepared from secondary xylem of P. tomentosa. Four anti-expression vectors with different fragments of PtoCesA1, named as pBIPF, pBICC1, pBIPR and pBIBR, were constructed. Some traits of transformed tobacco of pBICC1, pBIPR and pBIBR differed from wild types, such as small leaves, “dwarf” phenotype and thinner xylem and fiber cell walls than wild plants consistent with a loss of cellulose. It indicated that the growth of transgenic tobacco was restrained by the expression of anti-PtoCesA1. Transgenic tobacco was obtained and the contents of cellulose and lignin were analyzed as well as the width and length of fiber cells, and xylem thickness for both transgenic and control plants. Transformed tobacco showed a different phenotype from control plants and it implied that PtoCesA1 was essential for the cellulose biosynthesis in poplar stems. [Supported by the Hi-Tech Research and Development Program of China (863) (2001AA244060 and 2003AA244020) and National Basic Research Program of China (973) (J1999016003)]     

Variation analysis of seed and seedling traits of cross combination progenies in Populus

Twenty-five species and hybrids in Populus were used as parents, and 26 cross combinations, including more than 5 000 seedlings, were obtained by artificial cross breeding. The length of infructescence, number of seeds per infructescence, thousand-seed weight, germination rate of seeds among these cross combinations were tested. The results indicated that the cross combinational effects were significant for these traits, and demonstrated that the length of infructescence, thousand-seed weight were positively affected by female parent. In addition, seedling height, diameter above ground, diameter at breast height (DBH) of 17 cross combination progenies were investigated. The analysis of mean and standard deviation of these three traits showed that seedling height, diameter above ground, DBH had extensive variation among combinations and individuals within combination. Variance analysis and estimate of heritability indicated that the three traits had wide variation and were controlled by heredity. It was feasible to select superior cross combinations and seedlings. Further more, the result of multiple comparison showed that P. deltoides ‘Lux’ × P. deltoides ‘D324’, P. ussuriensis cl. ‘U4’ × P. deltoides ‘T66’, P. ussuriensis cl. ‘U4’ × P. deltoides ‘T26’, P. deltoides ‘Lux’ × P. ussuriensis cl. ‘U3’, (P. tomentosa × P. bolleana) × (P. alba × P. glandulosa), (P. alba × P. tomentosa) × (P. alba × P. glandulosa), and (P. alba × P. glandulosa ‘No. 2’) × P. tomentosa ‘Lumao 50’ were superior cross combinations with higher growth rate. Finally, 123 elite seedlings were selected for further test. [Supported by the Doctoral Foundation of Ministry of Education (Grant No. 200220022004) and the National “863” Project (Grant No. 2002AA241071)]     

Changes in water distribution in heartwood along stem axes in <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

 Three-dimensional distribution of water in the heartwood of Cryptomeria japonica D. Don was observed by soft X-ray photography. The within-tree variation in the distribution of “wet areas” (water-accumulated areas in heartwood) was enormous, as was the variation among trees. Although we found no universal pattern of changes in wet area distributions along the stem axes of all trees, similarities among individual trees within each cultivar and clone were observed. The difference between the two kinds of wetwood in C. japonica – genetically defined wetwood and secondarily induced wetwood – is discussed, as is the use of soft X-ray photography in Japanese tree breeding programs. Received: December 21, 2001 / Accepted: May 1, 2002 Part of this paper was presented at the 46th Annual Meeting of the Japan Wood Research Society, Kumamoto, Japan, April 1996 Correspondence to:R. Nakada     

Genetic diversity within and among southern African provenances of <Emphasis Type="Italic">Uapaca kirkiana</Emphasis> Müell. Årg using morphological and AFLP markers

Domestication of Uapaca kirkiana Müell. Arg is a high priority for improving rural livelihoods of smallholder farmers in southern Africa. Domestication efforts require knowledge of adaptive traits and intra-specific variation. Morphological traits and amplified fragment length polymorphic (AFLP) markers were used to assess genetic variation in twelve provenances of U. kirkiana collected from southern Africa. Assessment of morphological traits showed significant differences (P < 0.05) between provenances. Provenances from Zimbabwe and Zambia showed faster growth than those from Malawi (except Phalombe) and Tanzania. Morphological traits exhibited strong genetic differentiation between provenances and within provenances. The degree of provenance differentiation of traits (Q _ST = 0.03–0.139, Q _ST mean = 0.092) was in the same range with that of AFLP loci (F _ST mean = 0.089) detected among provenances and within provenances (0.002 < F _ST < 0.259). The differentiation for some morphological traits could be attributed to local adaptation and human selection of the U. kirkiana trees at the site of origin. Mean Nei’s (H) genetic diversity of AFLP showed high diversity within the provenances (H = 0.181–0.321, H mean = 0.256). An analysis of molecular variance (AMOVA) revealed higher genetic variation (90.8%) within provenances than among provenances (9.2%). There was no geographical pattern of variation in growth and morphological traits among the seed sources. Chipata provenance from Zambia was the most diverse while Mapanzure from Zimbabwe was the least diverse, though it was superior in height and earliest in fruiting. The pattern of genetic diversity indicates low selection in some areas and high gene flow which would be counteracting it. Therefore regional and country collections and conservation strategies should consider differences by focussing on the main range of the species by paying particular attention to unique populations within countries.     

Isolation and characterization of 10 new microsatellite loci in the ambrosia beetle <Emphasis Type="Italic">Platypus quercivorus</Emphasis>

The ambrosia beetle, Platypus quercivorus, is an insect vector for the causal agent of Japanese oak wilt disease, Raffaelea quercivora. We have developed 10 new polymorphic microsatellite markers from Type 1, one of the genetic types of the beetle, using a biotin-enrichment procedure. Fifty-three adult beetles were used for characterization of the markers. The number of alleles per locus ranged from two to nine (average 4.9). The observed and expected heterozygosity ranged from 0.49 to 0.77 and from 0.44 to 0.80, respectively. None of the loci deviated significantly from Hardy–Weinberg equilibrium. No linkage disequilibrium was detected in any pairwise comparisons of loci. Most of the markers successfully amplified in other genetic types of the beetle. These markers could become useful tools for population genetic studies and for parentage and sibship analysis for this beetle.     

泡桐基因文库的构建及actin基因同源顺序的亚克隆

以λ系列EMBL_3DNA为载体,LE392为宿主菌,建立了木本植物毛泡桐(Paulownia tomentosa (Thunb)Steudel)丛枝病抗病品系C161基因文库,命名为[λE_s/PTS(Bam-Sau)]。其包装效价达到1.1×10~6pfu,重组率为98%。用动物肌动蛋白(actin)基因为探针,从这一基因库中筛选出3.9Kb的泡桐DNA同源顺序进行亚克隆。所得结果表明:动物actin基因在木本植物基因组中存在一定的同源性。