A Greenhouse Survey of Resistance to Frenching in Nicotiana Species

Robert A. Steinberg Crops Research Division, Agricultural Research Service, USDA Beksville, Maryland, U.S.A. Tobacco Science, 1960, 4-47, p. 219-222, ISSN.0082-4523.pdf

The data dealt with herein are on frenching b.efore removal of the in- L,evels of abnormalities were re- frenchinp, a physiological disorder florescence would be satisfactory for corded as none, transient or perma- charact.eristic of, but not confined to, commercial tobacco production, since nent reticular chlorosis, and strapped Nicoticrnn (Karraker and Bortner, all new growth, i.e. all branches, are or small. Strapping had reference to 1934; Wolf, 1935). Th.e relation, if discarded. non-expansion of chlorotic leaf any, of frenching susceptibility to lamina without cessation of midrib of Nicotiftna species is un- Experimental Methods growth. Overall leaf proportions known. The disorder appears to be The techniques employed in this were almost normal in the “small” caused by Lalloiso-leucine, a toxin study of frenching responses hav,e lype of leaf response though size was liberated by Bctcillm cerelts Frank- been previously reported (Steinberg, greatly reduced. Abnormalities in land & Frankland, a soil saphrophyte 1956). Frenchlng capacity was pro- “small” type of response included (Steinberg 1951, 19523. Soil amend- duced in the soil by liming to neu- dorsal and ventral cupping and other ments, according to Clark and Smith trality and adding a large excess of departures in leaf shape exclusive of (1949)) caused wide variations in dicalcium ph0sphat.e after steam length x width relationship. Re- counts of B. census and other spore- sterilization. Nitrogen was kept to sponses were also recorded with re- fo’rming bacteria. Symptoms of a minimum. Frenching o’f ap- spect to minimum average time re- frenching begin as a network chlori- peared sporadically in the second quired for symptoms to appear in a sis in immature apical and crop and always became general in test lot and as the minimum for any may in severe cases include subse- the t.hird successive crop of tobacco single in all tests. quent non-expansion of leaf lamina, seedlings grown in the treated soil. retardation of stem growth, and All experiments were carried out in Experimental Results sterilitv. Previous determinations of the greenhouse at an air temperat,ure The responses among the Nicoti- susceptibility and immunity ob a of about 25” C. and a soil tempera- nna species are summarized in Table limited number of .Vicotinm species ture of about 35” C. The species des- 1. The minimum average time for were published by Wolf (1935) and ignations and authorities are those drenching in any one test is shown Sp.encer (19353. Agreement in re- considered valid by Goodspeed in th.e second column and the overall sults was poor. (1954). individual minimum in all tests in The present. paper includes a sys- Seed was germinated and the seed- the third column. Severi1t.y of leaf tematic surrey of 57 of the 60 recog- lings transplanted to 2-inch pots in symptoms ranged from none to nized species of Nicotiana (a single the usual manner. Seedlings of suit- strapped- or small-leaf formation. variety of each) with respect to de- able size for transfer (2-inch pots) The absence of responses by hT. ncau- gree of susceptibility to frenching were transplanted to the heat,ed beds. lia and N. thgkflora n.eeds verifica- and of differences in morphological Size of lots varied from 6 to 48, and tion. The very slow growth of both response. Teshs included a limited the tests were repeated as often as speci,es may have prevented appear- number of autopolyploid and inter- necessary. Each of the 4 frenching ance of typical sympto,ms. An atypi. specific crosses and a few species in beds was about 17.5 x 5.0 x 1.25 f,eet cal marginal chlorosis of apical other genera also reported to be sus- in size and could accommodate 96 leaves, however, was noted at t.imes cepti,ble. Among these was plants. Seedlings were usually grown in N. acnulis. nocturmin. L., of interest because of to seed formation unless stalk elon- All other available species were the postulated origin of Nicotiana as gation was suppressed by frenching subject to, frenching. Most responded a natural cross between progenitors and were then cut back to observe rapidly and in maximum degree. The of Petunia and Cestmm (Good- new growth. Symptoms of fr,enching only species showing any apparent speed, 1954). Resistant species might were generally intensified in the new degree of resistance were N. attentu- be suitable as genetic sources of re- growth. Tolerance as shown by nor- ata., N. f?.agrans, and N. petunioides. sistance to frenching for N. taba,cum. mal growth of new branches devel- No leaf strapping or other deforma- Resistance sufficient to prevent oped only after very long intervals. tion was noted in thes,e 3 species.

(Tobacco Science 219) reticular chlorosis. Table 1. Susceptibility to frenchinq in Nieotiana The reason for the 2 types of leaf Minimum time Type of abnormality when plants were af- Nicotiana species for frenching leaf fected most severely is unknown. The difference is not. considered to be ac- Aver.* Single Pk. symptomt cidental since it was verified several Days Days t.imes. This interpretation is support- ed by responses obtained with spe- acaulis s cies considered identical by some acuminata 29 18 n taxonomists (Goo’dspeed and Thomp- alata 18 14 s son, 1959). Small-leaf responses were ameghinoi given by (1) N. bigelovii and N. arentsii ib ii S clevelnndii and (2) by N. knightinna, attenuata 21 18 1’ but not by X. pnn~icztlnfa. Strap-leaf benavidesii 16 11 D responses were given by (3) N. longi- benthamiana 20 S flora and W. plumbnginifolia; by (4) bigelovii 1": 13 D LV. nesophilrr, N. wpandn, and N. bonariensis 26 21 S stocktonii; (5) by N. co?ymbosn. and clevelandii 30 21 D Tobacco Science, 1960, 4-47, p. 219-222, ISSN.0082-4523.pdf N. linearis; and (6) by N. pa.lmeG cordifolia 28 15 D and N. trigonophylla. A divergency corymbosa 32 25 S was found, th,erefore, in only one od debnevi 17 15 S these 6 groups of closely similar excelsior 19 10 S species. exigua 23 16 S Small leaf responses occurred in 5 f orgetiana 13 11 s of the 14 sections proposed by Good- fragrans 41 26 1’ speed (1954). Three (N. knightiana, glauca 11 11 N. bennvidesii, N. cordifolia) are in- glutinosa 16 10 : cluded in Section Paniculatae; N. goodspeedii 34 tom.en.tosn, in Section Tomen.tosae; gossei 28 2 E N. acuminntn, in Section Acumina- ingulba 19 14 S tae; N. bigelovii and N. cleTlelandii, knightiana 21 17 D in Section Bigelovinnae and N. mega- langsdorfii 14 14 S 10sip?107a,in Section Suaveolentes. linearis 18 10 S Species in which frenching took longibracteata . . place were not limit.ed to any part of longiflora is is S the range over which Nicotinna is maritima 33 24 S native. megalos,iphon 20 14 D miersii 64 34 S Table 2 contains a tabulation of nesophila 14 9 S the time element of frenching re- noctiflora 19 10 S sponses obtained with a few auto- nudicaulis 15 10 S polyploids and interspecific hybrids occidentalis 21 10 S of Nicotinna and of representativ,e otophora 16 14 S plants of 4 miscellaneous s#pecies. palmeri I6 10 S While all autopolyploids and inter- panicuIata 25 11 S specific hybrids tested frenched, a pauciflora 30 21 S few auto polyploids seemed to do so petunioides 21 16 1* less rapidly than the 2 n-species. plumbaginifolia 14 9 S These wel-e 4n-N. debneyi, 4n-hr. yos- raimondii 19 19 S sei, and 4n-N. occidentnlis. The dif- repanda 16 14 fer.ences found were small. rotundifolia 17 15 ss The 4 miscellaneous plants were rustica** 15 13 S representatives from 4 genera od setchellii 18 14 S Solannceae other than Nicotiann. All solanifolia. . 34 23 except Cestrum nocturnum L. spegazzlmi 29 28 ss frenched. Symptoms of ab’normality stenocarpa . . were usually mild in pepper and se- stocktonii 24 ii S vere in and petunia. Leaf suaveolens 23 10 strapping occurred in a single pepper sylvestris 17 13 : plant and in 2 in the new tabacum** 11 9 S growth folbwing decapitation after thyrsiflora N frenching in tests with 48 plant lots. tomentosa 11 11 D Many petunia plan& formed strapped tomentosiformis 19 14 S leaves in a similar experiment, while trigonophylla 21 21 S leaf shape and size remained normal undulata 25 15 S in Cestrum. velutina 21 16 S wigandioides 12 11 S Discussion The frenching response seems to b,e characteristic of Nicotiana and -

(Tobacco Science 220) Table 2. Susceptibility to frenching of autopolyploids and interspecific hybrids of Nicotiana and of miscellaneous plants Minimum time Minimum time Nicotiana autopoly- for frenching Interspecific hy- for frenching ploids and misc. Single brids of Nicotiana Single species Source* Aver. plant Source** Aver. plant

Days Days Days Days Nicotianrt 4rz bigelovii x tabacum 4 22 15 autopolyploids 412debneyi x clevelandii 5 24 21 4n alata 2 27 19 4n d.ebneyi x tabacum 2 18 7 4n benthamiana 2 27 18 412glauca x tabacum 2 18 11 4n debneyi 2 32 21 4n glutinosa x tabacum 2 22 17 472glutinosa 2 18 17 4n longiflora x tabacum 2 32 15 4n gossei 2 32 21 472suaveolens x tabacum 2 23 17 4n langsdorfii 2 15 11 4rz tabacum x cordifolia 6 16 10

4n longiflora 2 12 8 4n tabacum x knightiana 6 17 14 Tobacco Science, 1960, 4-47, p. 219-222, ISSN.0082-4523.pdf 4n megalosiphon 2 18 12 4n tabacum x longiflo’ra 3 18 14 4n occidentalis 2 32 20 4n undulata x tabacum 2 23 17 4n plumblaginifolia 2 14 7 4n rustica”” 2 20 18 2n tabacum x cordifolia 6 17 11 4n suaveolens 2 21 15 2n tabacum x knightiana 6 15 14 4n sylvestris 2 25 18 2n tabacum x wigandioides 6 20 11 4n tabacum 2 17 13 2n fragrans x tomentosifosmis 1 28 27 4n tabacum x 2n fragrans 1 16 13 Miscellnneous species Petunia . . 24 21 Pepper . . 38 35 Eggplant . . 17 14 Cestrum nocturnum . . Normal

* 1, L. G. Burk; 2, D. R. C~ncro~z; 3, E. E. Claytuu; 4, A. E. Kehr; 5, H. H. Snzith; 6, wviter. t With lots of xot less thnn 6 plants l * Variety brasilia.

perhaps other genera in the family for testing. Strong positive responses propriate conditions. . No correlations were were obtained in all genera of Wh,ether also the moderate resist- noted, however, between fr,enching Solannceae tested-tomato, pepper, ance to frenching found in certain capacity and taxonomic interrelation petunia, eggplant-except Cestrum. 2n and 4n species will prove serv- or habitat of Nicotiana species. Identification and utilization of a iceable for crossing with tobacco to Fifty-seven od 60 recognized species suitable source of resistance to increase its resistance will have to be elf Nicotiana w.ere tested and only 2 frenching may, therefore, prove dif- ascertainBed. However, even a small failed to give positive responses. ficult. increase in resistance to frenching in These were N. ncaulis and N. thyrsi- A frenching response in genera tobacco might prove adequate if flora, both perennials from South not in Solanacecre (ragweed, sorr,el, leaves on the stalk remain normal. America. These 2 species and N. and squash) has been reported by Frenching in axillary growth subse- ameghinoi are probably the only Jones (1953). Mild symptoms some- quent to cemoval of floral parts is winter-hardy perennials in Nicoti- what similar to t.he reticular chlorosis generally of no economic importance ana. The possibility exists that im- of frenching were also obtained by to growers. munity from frenching may be the writer in ragweed, cocklebur, linked 60 winter hardiness. Mildness plaintain, peppergrass, and dock. Summary of symptoms would indicate that Monomcotyledonous plant,s-corn, bar- Resistance to a physiological dis- N. attenuata, N. fra.grans, and N. ley-gave negative results. Verifica- order, frenching, was measured in petunioides may possess some resist- tion of these results would seem 57 of the 60 species of Nicotiana. ance. Habitats of these forms are desirable, since toxicity due to thal- There was no correlation between North America, Aust.ralia, and South lium (McMurtrey, 19321, manganese taxonomy o’r habitat and frenching America, respectively. Slowness in (Middleburg, 1954), lead (David, of species. Fifty-five of 57 species ap#pearance od symptoms would indi- Wark, and Mandryk, 1955), and per- tested gave a frenching res’ponse. cate th’at *either N. miersii from haps other elements may also c,ause N. acnulis S,peg. and N. thyrsiflora South America, N. stenocarpa from reticular chlorosis. Eitt. ex Goodsp. gave, no frenching Australia, or N. fragrans fro’m a The failure of Cestrum noctwnum response and N. a.ttenuata Torr., N. South Pacific islland might be best to give a frenching response is oif fragrans Hook, and N. petunioides in this respect. The species enu- interest since Nicotiana is considered (Griseb) Millan showed mild and merated are not limited as a group by Goodspeed (1954) to have origi- transient symptoms. Lag in appear- in range of habitat. Rang.e in resist- nated as a cross between progenitors ance of symptoms was greater in ance within each species is unknown of Cestrum and Petunia. Petunia N. miersii Remy, N. stenocarpa because of limited material availab,le frenched rather severely under ap- Wheeler, and N. fra,grans Hook. Only

(Tobacco Science 2.21) moderate increases in resistance rence of spore-forming bacteria on ence, N. 5’. 76: 86 (1932). were shown by a few autopolyploids plant roots and in soil.” 41st 4n- Middleburg, H. A. “Tobacco french- and interspecific hybrids. Other Sola- nual Meeting of Amer. Sot. Agron. ing and manganese injury.” Intern. naceac--tomato, eggplant, pew=- and Soil Sci. Sot. Amer., Milwau- Congress Soil Sci. Transactions. 5: and petunia-gave extreme french- kee, Wise. (1949). 2’78-283 (1954). ing responses though Cest,um noc- David, D. J., Wark, D. C. and Man- Spencer, E. L. “Studies on frenching turnurn L. remained normal. Nicoti- dryk, M. “L’ead toxicity in tobacco of tobacco.” Phytoputh. 25: 1067- ana has been considered to have resembles an early symptom of 1084 (1935). originated as a cross between pro- frenching.” J. Au&alian Inst. Steinberg, R. A. “Occurrence o’f Baci- genitors of Cestrurn and Petzlnia. Agr.Sci. 21:182-185 (1955). lus cer.e?ts in Maryland soils with The identity of symptoms in certain Goodspeed, T. H., “The Genus Nicoti- french.ed tobacco.” Plant Physiol. weeds classified in other plant fami- ana.” 536 pp. Chronica Botanica 26: 807-811 (1951). lies with those of mild frenching is Co., Waltham, Mass. (1954). . “Frenching symptoms pro- questioned pending verification with and Thompson, Mildred C. duced in Nicotiana tabacum and L-alloiso’leucine, the presumed toxin “Cytotaxonomy of Nicotiana II.” N. rustica with optical isomers of of Bacillus cpreus Frankland & Bo. Rev. 25:385-415 (1959). isoleucine and leucine and with Frankland. Species thought by some Jones, L. H. “Frenching of ragweed Bacillus cereus toxin.” Plant to be identical taxonomically, except (Ambrosia avtemisifolia L.).” Physiol. 27 : 302-308 (1952). N. panicula.ta L. and N. knightka Plant Physiol. 28:123-126 (1953). . “Prolduction and preven- Goodsp., agreed in showing either Karraker, P. E. and Bortner, C. E. tion o,f frenching o’f tobacco grown Tobacco Science, 1960, 4-47, p. 219-222, ISSN.0082-4523.pdf the small or the strap-leave response. “Studies of frenching of to’bacco.” in the green house.” Plunt and Kv. Agr. Exp. Sta. Res. Bull. No. Soil 7: 281-289 (1956). 349, pp. 63-109 (1934). Wolf, F. A. “Tobacco diseases and Literature Cited McMurtrey, J. E. “Effect of thallium decays.” 454 pp., Durham, N. C. Clark, F. E. and Smith, D. H. “Occur- on growth of tobacco plants.” Sci- (1935).

l

(Tobacco Science 222,)