(8.1)--植物检疫ISPM第27号附件19-Sorghumhalepense.pdf

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1、Produced by the Secretariat of the International Plant Protection Convention(IPPC)ENGDIAGNOSTIC PROTOCOLSINTERNATIONAL STANDARD FOR PHYTOSANITARY MEASURESISPM 27ANNEX 19DP 19:Sorghum halepense27 FAO encourages the use,reproduction and dissemination of material in this information product.Except wher

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4、ww.fao.org/contact-us/licence-request or addressed to copyrightfao.org.FAO information products are available on the FAO website(www.fao.org/publications)and can be purchased through publications-salesfao.org.The designations employed and the presentation of material in this information product do n

5、ot imply the expression of any opinion whatsoever on the part of the Food and Agriculture Organization of the United Nations(FAO)concerning the legal or development status of any country,territory,city or area or of its authorities,or concerning the delimitation of its frontiers or boundaries.The me

6、ntion of specific companies or products of manufacturers,whether or not these have been patented,does not imply that these have been endorsed or recommended by FAO in preference to others of a similar nature that are not mentioned.The views expressed in this information product are those of the auth

7、or(s)and do not necessarily reflect the views or policies of FAO.This diagnostic protocol was adopted by the Standards Committee on behalf of the Commission on Phytosanitary Measures in January 2017.The annex is a prescriptive part of ISPM 27.International Plant Protection Convention DP 19-1 ISPM 27

8、 Diagnostic protocols for regulated pests DP 19:Sorghum halepense Adopted 2017;published 2017 CONTENTS 1.Pest Information.22.Taxonomic Information.23.Detection.23.1 Preparation of samples for laboratory analysis.2 3.2 Sieve detection.3 4.Identification.34.1 Morphological identification of seeds.3 4.

9、1.1 Key to the seed morphology of Sorghum halepense and five related species.5 4.2 Molecular identification of seeds.5 4.2.1 Methods based on DNA markers.6 4.2.2 Controls for molecular tests.7 4.3 Biochemical identification of seeds.8 4.4 Morphological identification of plants.8 4.4.1 Key to the mor

10、phological characters of vegetative organs of Sorghum halepense and five related species.9 4.4.2 Key to the morphological characters of reproductive organs of Sorghum halepense and five related species.9 4.5 Cytological identification of plants.10 4.5.1 Chromosome counts.10 4.5.2 Flow cytometry.10 4

11、.6 Comparison of the confidence level of the identification methods.11 5.Records.126.Contact Points for Further Information.127.Acknowledgements.128.References.139.Figures.15DP 19 Diagnostic protocols for regulated pests DP 19-2 International Plant Protection Convention 1.Pest Information Sorghum ha

12、lepense(Johnsongrass)is a perennial grass with a ribbed leaf sheath,conspicuous midrib,large,purplish panicles,and far-reaching rhizomes(Figures 1 and 2).Its origin remains unclear,but some authors suggest that it originated from the hybridization of Sorghum arundinaceum and Sorghum propinquum throu

13、gh chromosome doubling(chromosomes:2n=4x=40)(Nguni et al.,2010).S.halepense is native to the Mediterranean area(Meredith,1955)and has been introduced to other regions(Bor,1960).It has become widespread,and is distributed from latitude 55 north to 45 south.It is best adapted to warm,humid areas with

14、summer rainfall,areas with a high water table,and irrigated fields in subtropical zones.S.halepense is one of the most malignant weeds worldwide,impacting more than 30 cereal,vegetable and fruit crops(Holm et al.,1977).It also threatens biodiversity in at least 50 countries in temperate and tropical

15、 areas throughout the world,including countries in which it is a native species(Holm et al.,1977).The main factors affecting the pest risk of S.halepense as a pest of plants are that:(1)it has a high asexual and sexual reproductive capacity;(2)its seeds can be dormant and are long-lived,and can move

16、 with traded commodities(Warwick and Black,1983);(3)it has strong competitive ability and causes great yield loss in crops(Follak and Essl,2012);(4)it is an alternate host of numerous pathogen species;(5)it has allelopathic effects and is toxic to livestock(da Nobrega et al.,2006);(6)it has develope

17、d resistance to a wide range of herbicide groups(Heap,n.d.);and(7)it has self-compatibility but readily crosses with related species,which may result in more invasive hybrids or cause gene introgression of crop species(Warwick and Black,1983;Arriola and Ellstrand,1996).S.halepense is able to reprodu

18、ce by rhizomes or seeds.Rhizomes readily sprout and can be distributed by tillage.An individual S.halepense plant is able to produce as many as 28 000 seeds in a growing season.These seeds are able to survive and germinate under most environmental conditions.The seeds are caryopses and are brown,obo

19、vate,3 1.6 mm in size,with an elliptic sessile spikelet that is appressed pubescent(Table 1).Seeds are the main means of spread of S.halepense,and they are readily distributed naturally by wind and water as well as by birds and other animals.More importantly,the seeds are frequently disseminated by

20、human activity as a contaminant of commodities traded around the world;in particular,crop seeds and raw grains,such as Sorghum bicolor(sorghum),Glycine max(soybean),Zea mays(maize),Triticum aestivum(wheat)and Sesamum indicum(sesame),as well as forage,Gossypium spp.(cotton)and birdseed mixes.2.Taxono

21、mic Information Name:Sorghum halepense(L.)Pers.,1805 Synonyms:Holcus halepensis L.,1753 Taxonomic position:Plantae,Angiospermae,Monocotyledonae,Poales,Poaceae Common names:Johnson grass,Johnsongrass(English)3.Detection Common survey methods for herbaceous species may be adopted for the detection of

22、S.halepense in the field.In order to detect seeds of S.halepense in crop seeds,an inspection procedure should be followed in which a composite sample is prepared for laboratory analysis and sieve detection(ISTA,2014).3.1 Preparation of samples for laboratory analysis General guidance on sampling met

23、hodologies is described in ISPM 31(Methodologies for sampling of consignments).The sample for examination should be approximately 1 kg.Remaining sample material Diagnostic protocols for regulated pests DP 19 International Plant Protection Convention DP 19-3 should be labelled and conserved in paper

24、bags or glassware free from moisture for possible further checking.3.2 Sieve detection A set of three sieves should be assembled with decreasing aperture sizes according to the seeds or grains being sampled,within an overall range of 2 mm to 10 mm.The largest aperture sieve is placed on top of the s

25、econd largest sieve,with the smallest sieve on the bottom.The sample for examination is placed in the top sieve and the sieve set assembly is covered before sieving the sample through it.After sieving,the material remaining in each sieve layer is collected and placed onto white plates for visual exa

26、mination.The suspected S.halepense seed fragments and seeds(resembling those shown in Figure 3)are selected for further identification.4.Identification Identification of S.halepense seeds is the main task and is commonly based on morphology.For suspected seeds with intact glumes and upper lemmas,mor

27、phological identification methods(section 4.1)are reliable.However,the fruits and seeds collected may be incomplete and parts of their characters unclear.In such cases,molecular(section 4.2)or biochemical(section 4.3)identification methods may need to be used.Seeds may also be sown and grown into se

28、edlings and mature plants,either of which can be morphologically(section 4.4)or cytologically(section 4.5)examined for taxonomic traits and subsequently identified as a complement.Figure 4 presents a flow chart for the identification of S.halepense.S.halepense is prone to be confused with five relat

29、ed species in the genus Sorghum:-S.almum Parodi(S.bicolor subsp.drummondii(Nees ex Steud.)de Wet ex Davidse),1943 -S.bicolor(L.)Moench,1794 -S.propinquum(Kunth)Hitchcock,1929 -Sorghum spp.hybrid cv.Silk(silk sorghum),a hybrid between Krish hybrid sorghum(S.halepense S.roxburghii)and S.arundinaceum,1

30、978(CSIRO,1978;Flora of China Editorial Committee,1997,2013;Ross,1999;Barkworth,2013).-S.sudanense(Piper)Stapf,1917.This diagnostic protocol compares S.halepense with the above five closely related species.Detailed descriptions of plant morphological characteristics can be found for S.halepense in H

31、olm et al.(1977)and Flora of China Editorial Committee(1997,2013);for S.almum,S.bicolor,S.propinquum and S.sudanense in Flora of China Editorial Committee(1997,2013);and for Sorghum spp.hybrid cv.Silk in CSIRO(1978)and Ross(1999).4.1 Morphological identification of seeds The caryopsis of S.halepense

32、 is brown,obovate,2.63.2 mm in length and 1.51.8 mm in width;obtuse in the apex with persistent style;hilum rotund,deep purplebrown;ventral side flat;embryo oval or obovate,with length approximately one-third to half of the caryopsis(Figures 2 and 3).S.halepense seeds can be identified based on char

33、acteristics of the glume and upper lemma(Tables 1 and 2).A key for species identification can be used to distinguish similar species if a seed is not easily matched to the description of characteristics in Tables 1 and 2.DP 19 Diagnostic protocols for regulated pests DP 19-4 International Plant Prot

34、ection Convention Table 1.Comparison of the sessile spikelet,caryopsis and seed weight in Sorghum halepense and five related species Species Sessile spikelet Caryopsis Weight of 1 000 seeds(g,approximate)S.halepense Elliptic or ovate,(3.8)45(6.5)mm in length,appressed pubescent Dark brown,obovate or

35、 elliptic,2.63.2 mm in length and 1.51.8 mm in width 4.9 S.almum Elliptic to oblong,4.56.5 mm in length,short pubescent Redbrown,broadly ovate or oval,3.34 mm in length and 22.3 mm in width 6.6 S.propinquum Ovate,or broadly ovate,3.85 mm in length,bearded Brown,broadly ovate or broadly oval,approxim

36、ately 2 mm in length and 1.5 mm in width 3.8 S.sudanense Elliptic,(5)68 mm in length,sparsely pubescent Redbrown,broadly ovate,3.54.5 mm in length,2.52.8 mm in width 1015 S.bicolor Elliptic to oblong or ovate,(3)4.5 6(10)mm in length,densely hispid,or pubescent to glabrous Pink to redbrown,ovate,3.5

37、4 mm in length,2.53 mm in width 20 Sorghum spp.hybrid cv.Silk Oval,approximately 3.8 mm in length,short pubescent Yellow or yellowbrown,broadly ovate,2.54 mm in length and 1.72.5 mm in width 4.2 Source:Based on Holm et al.(1977),Sun et al.(2002),Qiang(2009),Barkworth(2013),Flora of China Editorial C

38、ommittee(2013)and Clayton et al(2016).Table 2.Comparison of the glume and upper lemma of seeds in Sorghum halepense and five related species Glume Lower glume Upper glume Upper lemma S.halepense Subleathery,tawny,redbrown or purpleblack Apex clearly tridenticulate,57-veined,dorsum ciliary but the re

39、st glabrous 3-veined Triangular lanceolate,apex bilobed and awned or not;awn 1016 mm S.almum Chartaceous or subleathery,dark brown Apex slightly tridenticulate,57-veined,dorsum ciliary but the rest glabrous 3-veined Lanceolate,apex obtuse or slightly acute,bilobed,awned;awn approximately 15 mm S.pro

40、pinquum Subleathery,dark brown with inconspicuous crossveins 911-veined,apex acute to apiculate or tridenticulate,pubescent 7-veined Lanceolate,approximately 3.5 mm in length,acute or emarginate,awnless Diagnostic protocols for regulated pests DP 19 International Plant Protection Convention DP 19-5

41、Glume Lower glume Upper glume Upper lemma S.sudanense Leathery,lemon yellow to redbrown Apex bidenticulate,1113-veined,usually with crossveins,dorsum short ciliary 57-veined,with crossveins Ovate or elliptic,apex bilobed,awned;awn 1016 mm S.bicolor Leathery,pink to redbrown Apex acute or tridenticul

42、ate,1216-veined with crossveins,dorsum dense ciliary 79-veined Lanceolate to long oval,24-veined,apex bilobed,awned;awn approximately 1 mm Sorghum spp.hybrid cv.Silk Leathery,tawny,redbrown or purpleblack Apex slightly tridenticulate,57-veined,dorsum ciliary but the rest pubescent 3-veined Broad lan

43、ceolate,apex slightly bilobed,awnless Source:Based on Holm et al.(1977),Sun et al.(2002),Qiang(2009),Barkworth(2013)and Flora of China Editorial Committee(2013).4.1.1 Key to the seed morphology of Sorghum halepense and five related species Based on Holm et al.(1977),Qiang(2009)and Flora of China Edi

44、torial Committee(2013).1.Glume with clear crossveins;lower glume with more than 11 veins;large seed weight(1 000-seed weight 10 g).2 Glume with no clear crossveins;lower glume with 11 or fewer veins;small seed weight(1 000-seed weight 615 nm is detected with a photomultiplier screened by a long pass

45、 filter.The mean 2C DNA content of each target species is calculated by comparing its mean nuclear fluorescence with the mean nuclear fluorescence of an 2 In this diagnostic protocol,methods(including reference to brand names)are described as published,as these defined the original level of sensitiv

46、ity,specificity and/or reproducibility achieved.The use of names of reagents,chemicals or equipment in these diagnostic protocols implies no approval of them to the exclusion of others that may also be suitable.This information is given for the convenience of users of this protocol and does not cons

47、titute an endorsement by the CPM of the chemical,reagent and/or equipment named.Laboratory procedures presented in the protocols may be adjusted to the standards of individual laboratories,provided that they are adequately validated.Diagnostic protocols for regulated pests DP 19 International Plant

48、Protection Convention DP 19-11 internal standard.Because of the variation of Sorghum DNA content,one of two different internal standards is used to avoid overlap of the standard and target species.One standard,Arabidopsis thaliana ecotype Columbia,has a genome size of 157 Mb or 1C=0.16 pg.The DNA co

49、ntent of A.thaliana and S.bicolor Tx623(2C DNA content=1.67 pg)is determined from 15 replicates of leaf samples from S.bicolor and A.thaliana Columbia.At least three replicates for each test sample are analysed to obtain the mean DNA content(Price et al.,2005;Jessup et al.,2012).Samples with more th

50、an 30 chromosomes can be suspected to be S.halepense,or S.almum or non-flowering Sorghum spp.hybrid cv.Silk.S.propinquum,S.sudanense,S.bicolor and flowering Sorghum spp.hybrid cv.Silk can be excluded.Furthermore,samples with 40 chromosomes can be identified as S.halepense when S.almum and non-flower