As one member of the largest transcription factors family,MYB gene plays an important role in the processes of second metabolisms and growth development in plants. A gene which was homologous with MYB was cloned from Indosasa hispida cv. ‘Rainbowu0027by the rapid amplification of c DNA ends( RACE) method,and named as Ih MYB4. The Ih MYB4 expression in different organs of I. hispida cv. ‘Rainbow u0027 was tested by RT-PCR. The sequence analysis showed that the c DNA sequence of Ih MYB4 consisted of 1044 bp encoding347 amino acid,and contained two Myb-DNA-binding domains,indicating that it was belonged to the R2R3-MYB family. Phylogenetic analysis,based on protein sequence,showed that the Ih MYB4 protein was likely to be involved in the regulation of anthocyanins biosynthesis. The Ih MYB4 was expressed only in young red culm by RT-PCR,suggesting that the Ih MYB4 participated in the regulation of anthocyanins biosynthesis of I. hispida cv. ‘Rainbowu0027.

Flavonoid-3-O-glucosyltransferase (3GT)is the key enzyme in anthocyanins biosynthesis,which trans-forms unstable anthocyanidins into stable anthocyanins.The function of Flavonoid-3-O-glucosyltransferase in bam-boo is unclear,although many flavonoid-3-O-glucosyltransferases from other plant had been reported,so far.In-dosasa hispida McClure cv.Raln bow is of few bamboo species which can produce anthocyanins in clum.Therefore,I .hispida McClure cv.Raln bow is very important material to reveal the function of Flavonoid-3-O-glucosyltrans-ferase in bamboo and the mechanism of anthocyanins biosynthesis in bamboo.Therefore,it is first step to clone 3GT gene from I.hispida McClure cv.Raln bow.First,gene special primers of 3GT were obtalned based on the homology analysis of reported flavonoid-3-O-glucosyltransferase,and the RNA was extracted from yound red clum which pro-duce anthocyanins part in I .hispida McClure cv.Raln bow by Trizol method.And then gene fragment of I .hispida McClure cv.Ralnbow was cloned with special primers of 3GT.According to the obtalned fragment sequence,the primers were designed,which were used in rapid amplification of cDNA ends (RACE).Next,the 3′end and 5′end sequence of 3GT were obtalned by RACE,the full length gene of Ih3GT was assembled by ATG software.Finally, the full length gene of Ih3GT was cloned from I .hispida McClure cv.Ralnbow by reverse transcription-polymerase chaln reaction (RT-PCR).ORF analysis program was used to confirm open reading frame,MEGA software was used to construct phylogenetic tree.the DNAman software was used in Homology analysis,and semi RT-PCR was applied in gene expression profile.The results showed that the cDNA sequence of Ih3GT consisted of 1 425 bp open reading frame (ORF)which encodes 474 amino acid,Ih3GT and 3GT from Poaceae were grouped in same clade,the deduced protein of 3GT from I.hispida McClure cv.Raln bow shared 69% identities with 3GT of Oryza sativa and shared 67% identities with 3GT of Brachypodiumdistachyon.Homology analysis showed that deduced Ih3GT protein had a glycosyltransferase signature domaln PSPG-box.Expression profiling with semi RT-PCR analysis revealed that Ih3GT was expressed in young red culm and was not expressed in old culm,old leaf,young leaf and shoot.This im-plied that the expression of Ih3GT from I.hispida McClure cv.Raln bow showed obvious tissue specificity.This study will provide useful information to reveal mechanism of anthocyanins biosynthesis in I .hispida McClure cv. Ralnbow in future.Ih3GT gene can be transformed into Eschera coli,and heterologous expression obtaln Ih3GT pro-tein and detect the enzyme activity of Ih3GT,and it can also be transformed intoArabidopsisthalianaorOryzasati-va,the function of Ih3GT will be confirmed by heterologous expression.The obtalned Ih3GT also can been implied in bamboo breeding or other horticultural plants by genetic engineering.

Agrobacterium tumefaciens-mediated transformation(ATMT) was successfully applied to the lichen forming fungus Cladonia metacorallifera. Vegetative mycelium of this fungus was used for transformation by ATMT. The genes for hygromycin resistance(hph) and the enhanced green fluorescent protein(EGFP), both under the control of a trp C promoter, were employed as selection markers. PCR amplification of hph and the observation of fluorescence signal by confocal microscopy showed that C. metacorallifera was successfully transformed by ATMT.

通过已报道查尔酮合成酶基因的保守序列设计特异引物,扩增得到七彩红竹(Indosasa hispida cv.rainbow)查尔酮合成酶(Chalcone Synthase,CHS)基因片段,再以RACE法获得CHS基因全长cDNA序列.该cDNA全长1953 bp,含1542 bp的开放阅读框,编码含513个氨基酸残基的蛋白质.多序列比对结果表明IhCHS1推断的蛋白与节节麦(Aegilops tauschii等禾本科植物的CHS相似性在87％以上,临位法构建系统发生树显示IhCHS1与禾本科植物CHS亲缘关系较近.RT-PCR结果显示IhCHS1在幼嫩红秆中大量表达.

Dihydrolfavonol 4-reductase (DFR) is a key enzyme in the anthocyanins biosynthesis pathway, and plays a critical role in flower pigmentation. The gene-specific primers were designed according to the transcriptome sequencing data, and the full-length cDNA of a novel DFR gene was cloned from Indosasa hispida cv.‘Rainbow’ with the method of reverse transcription PCR. This novel gene was named as IhDFR1 (GenBank accession No. KF728205). Sequence analysis indicated that IhDFR1 was 945 bp in length and encoded a protein with 314 amino acids. Bioinformatics analysis showed that IhDFR1 had the typical functional domains of DFR protein, containing two speciifc binding sites and belonging to the non-Asn/Asp DFR. The IhDFR1 was homologous with the DFRs from the gramineous plants. The temporal-spatial expression analysis based on different growth stages indicated that the IhDFR1 was expressed only in the reddish violet culms. The results above preliminarily suggested that the IhDFR1 might be an important enzyme governing anthocyanin metabolism, and lay a theoretical basis for further exploration of molecular mechanism of anthocyanins and for the comprehensive exploitation and utilization of I. hispida cv.‘Rainbow’.

Chalcone isomerase ( CHI) is one of the important structure genes in Anthocyanins biosynthesis.IhCHI1 was successfully obtained from Indosasa hispida cv.‘Rainbow ’ by RT-PCR with special primers designed based on the transcriptome data , and this sequence accession number is KJ 477333 in GenBank.Sequence analysis showed that IhCHI1 had complete cDNA open reading frame ( ORF ) , which consisted of 690 bp encoding 229 amino acids.Blast analysis showed that IhCHI1 belonged to the CHI family.Phylogenetic analysis showed that Indosasa hispida cv.‘Rainbow’ had close relative rationship with Oryza sativa and Zea mays.IhCHI1 expression analysis in different organs of Indosasa hispida cv.‘Rainbow ’ by RT-PCR showed that IhCHI1 strongly expressed in young red culm.

采用L9(34)正交试验设计,了解发芽环境、微波辐射和激素浸种对云南松种子发芽的影响,结果表明,3种因素共同影响云南松种子的发芽。其中,发芽环境是影响发芽率和发芽势的主要因子,夜间无光照的温度相对较低环境适宜云南松种子的发芽,但与全天光照且温度相对较高环境的差异并不显著。微波辐射对云南松种子发芽的影响比激素浸种小,2个因素均显著地影响云南松种子的发芽率,但都对发芽势的影响不显著。发芽率和发芽势最佳的组合分别为A3B1C1和A3B2C1,即无微波辐射、清水浸种置于夜间无光照环境下的种子,和微波辐射15 s、清水浸种置于夜间无光照环境的种子,其2个指标分别达83.0%和48.8%。

The L9(34) orthogonal design was applied to understand the effects of germinative environment,microwave radiation and hormone on radicle and hypocotyl length of Pinus yunnanensis seedlings.The results show that optimal treatment combination of improving radicle and hypocotyl length was whole-day lighting,non-microwave radiating and 0.2 g/L solution of IBA soaking seeds.Germinative environment significantly influenced radicle and hypocotyl length,in which whole-day lighting was benefit to elongate radicle and hypocotyl(19.88mm and 45.69mm,respectively).Hormone solution soaking seeds primary affected on radicle length.0.2 g/L concentration of IBA solution facilitated radicle growth.There were no significant differences on above 2 parameters between the levels of microwave radiation.