- ______,.. ___,______,,_._~,~~ ~~-==~~·~~---.--.~--~

STUDIES I IT PLlllTT HISTOLOGY,

CYTOLOGY, !1ND SYSTE!.1.!~ TIC BOT.t\ITY.

UAJOR ~HESIS IN BOTANY .

presented by

F.

for the dogreo of Master of Science.

VIRG I 1TI ...I\. POLYTECIIlTIC I1!STITlTTE .

Jtme 1, 1911.

-

. .. .. ,_ -

-·

• • • ----STUDIES -IN PL.A.NT--- HISTOLOGY,------CYTOLOGY, .1.ND SYSTELil1TIC BOTANY.

INTRODUCTION.

This paper is designed to give an account of the work done by the writer in a oourse of Plant Histology and Systematic Botany for the degreo of llastcr of Science. The l!laterial has been. gathered from a num- ber of sources. Besidoa the booka ·named in tho bibliography, which is found at the end of this pnper, various periodicals such as"Science"and the "Botanical Gazette" have been consulted in looking up special points. This discussion naturally falls under three heads: first,

~.~iorosoopic Teohnique; oocond, Histology and Cytology, and tl1ird, Syster.iatic

B otany~ Tho first two divisions are very closely related and might well be considered together but for convenience of treatnent and olcurness, it is thought best to separato them. #2 • •

PART I.

MICROSCOPIC TECHNIQUE.

While it is not possible in a discussion of this length to ·go into details in describing the various steps used in the histological laboratory, tho more important points will be briefly noticed. It may be said, however, that all point.a are of prime importance in the laboratory.

The overlookins of a single apparently uni~portnnt detail ~ay be the cause of absolute failure. Carefulness and ·o1eanliness must be the con- stant watoh,vords of those w:ho would succeed. A description of the various pieces of apparat'l1a is unnecessary and will not be attempted. The one most important piece of apparatus- if there is such a thing- is the compound microscope. In fact, we can not oonoeive of histology or oytoloGY without the compound microscope. The first compound microsoope was r-ade about 1600 , and it has been continually improved sinoe that time. ~he improvement of the microscope han l~ept po.oe with the work in histology and cytology. The microscope reveals a field as rich and varied ao tre most enthusiastic investigator could desire, and the re sult ·a of the last half cent1iry show that the field has not been neglected.

~IBTHOD USED.

~he paraffin method was used in ~ost of the work done, end the discussion of the various steps ta1ron applies to this oet11od. It is the most used of all the methods of the histological laboratory. By the use of the rotary rniorotoma, or even the slide microtome·, very satisf)actory work can be done. •

KILLING A]ID FllI1TG.

The first step in the preparation of material is the killing. The object of killing i' to bring to a sudden and complete stop, the life process. By doing this, we can observe the cells as they actually were at the instant they v1ore lrilled. One other step is necessary, l1owo11er, if we are to see the result of the killing, that is fixing. The fixing so hardens the cello that they retain their original condition .through all the subsequent steps. With animals, and sometimes with plants, one agent is used for lrilling and anothe1.. for fixing. l1B a ri1le, only one agent is used for botl1 processos with plants, since they have r. o foe ling in the sense that animals have and this is less troublesome.

A number of ~gents aro en:ployed; alcohol, chloroform, formalin, c'orrosive, sublimate, and various ncido aro the r..iost c o~on. A stock solution of the following formula;

Chromic acid c.rystala 10 g. Glacial acetic acid 10 o.o. Water 1000 o.o. was kept for diluting to the proper strength as needed. The amount of dilution necesnary must be deter~ined in each case by experiment. It may be desirable to change the proportion of chromic and acetic acids. Chromic acid causes contraction and acetic acid causes ·swelling. Anothar formula is;

(1 peroent chronic acid 250.0. A-(1 percent acetic acid lOo.o. (Water 55c.c.

B- 1 percent oa~ic acid lOc.c. ... The osmio acid io added to the mixture A as it is needed for use , s i nee the +vWO d o n o t 1reep vre 11. tope+her· · o · v · • It gives good results where ... II·.i4

one wants to study·mitosio. Flening's triple stain gives its beat results

when used after it. The should bo removed from tho nato1·ial and it

should be cfit into o~all piecoo before it is placed in the killing fluid, so that it will bo penetrated quickly.

· The material should romain in the fixing flu~d fro~ six to twenty four hours according to the ohnractar of the tissue.

WASIII~IG.

The next step in this process is the re~oval of tho killing and fixing agent which if left would interfere with subsequent stepo or leave

precipitations w11ich wo11ld bo rni~leo.ding. The best v1ay to do this is to leave the matorial in a vesoel under a slowly running faucet of water over night.

HARDE.?TING .AND DElIYDR.'lTING.

Tho fixing agent, as already stated, begins the hardonins but this must be oontinuod as soon ao the washing is comploted, end since the material must also be dehydrated, it is passed thro1lgh a series of alcbhols

of different strengths. From tho water, the ~aterial should go to 15~ .• where it should remain from 6 to 24 hours. It should then be loft for the same length of time sucoossivel~T in 35 %, 50~0 , 70%, - 85%, 95%, and 100% alcohol. This finishes the hardening and the dehydrating proce3s. As

far as hardening ie concorne~,the 70% alsohol perhap~ hardens the ~ate­ rial sufficiently but it must go on through the 100% alcohol to co~plote the dehydration. CLEARIJTG.

After the material is completely dehydrated, the alcohol must be

This is done by the uao of sylol. Other agonto may be used, but

this is most o cmunonly employed and perhaps, it is the best. Lilre the :/1 5 11 • hardening and dehydrating procesoos, the change from alcohol to ~lol should be gradual. From the 100% alcohol the material shonld roraain from 6 to 12 hours successively in 3 parts 100% alcohol and 1 part xylol, 2 parts 100% alcohol and 2 parts xylol, 1 part 100% alcohol and 3 parts xylol. The material should now go to pure xylol where it should remain till it is clonr. 'Thio time will vary greatly with the material.

INFILTR~\TION.

The embedding also is n gradual process. Tho first step is to place a pieco of paraffin in the bottle of zylol. Thia should not be larger than tho xylol will diosolve. The bottle containing the xylol paraffin mixturo should bo kept wnrrn over night. From horo tho material goes to soft paraf fin ( 43° C. ) for from 2 to 12 hours, depending upon the oharucto~ of tho tisoue and its size, from which it is put into the harder paraffin (52° C.) for the name or a greater length of time. Tho temperattrre of the bath io l:opt abot1t 1 ° above this at 53° C.

filiBEDJJING.

Tha material is now ready to imbed -in paraffin in which it may in~efinitoly. The harder grade (52° C.) is used for thio purpose.

Paper trays of the aize requirod by tho amount of material to be imbedded Phoy should be placed in a tray with wax in the bottom near the temperature of the paraffin. A layer of paraffin, deep enough to coyer material io now poured into tho tray along with the material or it be put in with a JE,ir of wnrrn forceps. A warm needle is useful in within the paraffin. Water should be poured around tlie paper trayo at once and, as soon as the paraff in hardens ouffic i ontly to bear the weight of !t, water should be poured on top of tho paraffin. If the paraff in cools slowly, it doos not nako good ribbons when it is cut. .,r.,l~ 6 I/

SECTIONI:tTG.

Cutting may begin as soon as the paraffin cools or it may be delayed any length of time desired. !f a good riobon of sections is to oe secured, we cust watch several. points. The microtome ~ust be in good order, the temperature must be· right, and the bloclr :rn11st be n perfect rectangle. The microtome should be well oiled and t~e knife sharp. The temperat11re of the room in '\v!1ich tho cutting is done should bo neither very hot nor vary oold. If the blook is too- warm, it ~ay be cooled by placing it in cola water for a shert while; if too ·cold, ones breath or thumb upen the knife a few minutes ~ay make the sections ribbon as de- sired. Unless the block is cut a perf ect rectangle and all points strike the knife sinultaneoualy, the ribbon will be crooked and this causes much waste of space on the slide as well as being very annoying. Practice is

j necessary to be able to mul{e good ribbons and even this sornetiI!lco fails. material is uaually cut betwcon 2 and 10 "If f' thick. FIXIITG TO SLIDE

Tho length of riobon or ribbons to be put on the slide will

upon the size. of the cover glass. The slide should be cleaned perfectly with alcohol, and thon a small drop of albumen fi~ativo should be spread evenly upon it with tho finger. The ribbon is now placed upon and a few drops of water placed around it. The wator ia now flame until the ribbon flattens out, whon the water around is wiped off and tho slide is left over night or for at least four hours in the trays of the paraff in bath when tho section should be found firmly attached to the slido. RE110V1.L -OF THE P }JtAFF IN.

Th·e paraffin is aomotimea removed fror.i the slide by heating it

till it rcaoheo the n1elting point and then washing it off with turpentino, but, sinoe this is apt to end diaastorously, the turpentine bath was used.

The turpentine re~ovcs the paraffin in a few minutes. RE ~1 0VAL -OF ~HE TURrENTilfE.

The slide is tal:en from the turpentine bath, and as much of

the turpentine as poesible removed by wiping tho slide with a cloth. Tho slide is n ow rinsed off thoroughly with 100% alcohol and thon placed in a Stender dish of 95 % alcohol. The rmterial is now ready for tho stain unleso it is a water stain in which case the olide should first be dipped in water in order not to carry any alcohol over into the otain •.

While it is true that there is yet much to be learnod of steins

and their 11ae . in histological and cytological worl~, it is true, ne"T1crthele ss

that tho n1unber of them is very large. Stains nay bo olaas~fied in nany ways: as general and opecific; acid and basic; and into throo groups; the Haematoxylina, the Analins, and the Carmines. Since evon a brief discus-

sion of stains would na1~e this pnpor too long, t11oy will not be further noticed except to explain the noat characteristic properties of thooo used

in the laboratory hero.

t 1loot of the oytologicul \70rlrers of recent yoaro ha"1e uoed one of

two stains in their worl~-, Haide~~uin's iron- hnomatoxylin, or Fle~ing's triple stain. Tho second, however, has been uoed exclusively in cytological . . ' . work, and a brief discuosion of it may properly. proceed nn account of the staining done with it w~ ich is to follow. As above stated, Flening's Safranin, Gentian- Violet, OrangeG . . oombinution was uaed for all mitotic fieuros. The stains are mode as fol-

lows: To a 1% solt1tion of alcoholic safranin ( 100 or 95~~ alcohol) a.dd an equal voltune of 1% aqueous solution of sa: 1ranin in distilled ,-,ater; a i·;b ~queous solution of oreange Gj l'J ~1 ~,~-~ The slide ~ay be placed in the safranin from any grade alcohol or water. In the case of tho parnf~ fin riethod, it of course, cor.ioo fr om the 95 ; alcohol used for washing away the tur~cntine. In the work here it was found that slideo placed in tho oafranin juot before leaving the laboratory in tho afternoon- about 5 o'clock- were stuinod nufficicntly by eight or nine o'clock the next norning. !t is best to leave tho ol~dc in the safranin till the ~ntorial is quite a bright red, and then reduce. urUt_ This is done by rinsing.A.50% alcohol, or slightly acidulated alcohol, until the proper dogroo of differentiation is reached. The chro~osonco nh ould be a clear, bright red. The slide ohould be places in water to remove all the alcohol and thon go to tho gentian violet for about 5 min11tco. '.!:he gentian violet is intended to ctain the spindlo and not tho chronooones. The slide should bo rinsed in uutor bcforo being placed !n the orange G. whera it should rocain about l oinuto. It should bo rorneobored that the orange G. reduces gentian violet. It stains the cytoplasm. Tho olide should then be dehydrated uith 95 and 100% alcohols, beinc careful not to t1se too mt1oh alool1ol or to nllow it to remain on the slide too long.

The matorial io then cloarod with clove oil and th~n io re~oved with xylol to prevent tho fading of the acrornatic figures. No dofinite rule can be given f or the t:.rie the nlido must be lef t in any of the otains.

The only way to obtain the best rooults io by repeatod experiments in ench particular caso. Before the naterial on the slido nan ti~e to dry, it is covered with baloan, and the cover glans which hao boen run through the flame of the Bunsen burner is placed c~er the ~aterial. ~ ith a little prac-tice, it is easy to j11dge jt1ot how much balsari to use so as . to f ill out the cover glaso co~plotcly and still not gu~ up the olide.

balsam hnrcions in a fe\11 do.:ro and the adc1 ition of o. label mal~co motmt c.ompletc.

The paraf fin ~ethod in long and tedious, but because of its

ad ·vantages no other hao yet been perf ected that can tal{e its pl£1cc for many 1rinds oil hiotolor;icul -r1orJr. , • PP.RT II •

.STRUC~URE OF THE CELJ;.

Any discussion of histology or cytology must be prefaced by a discussion of the structure of the cell. Cells are the elernontary organo of animals and plants. Unless otherwise mentioned, tho cell of the flowering plants will be referrod to herein.

1legotn.ble colls Tiere first noticed by Robert Hoolro in bottle cork in the year 1667. Tho true character of the coll was not known till about the middle of the past century. Since that time, a host of investi- gators have added to our knowledge of tho structure and functions of t h e cell. The wo~ . cell is not very well chooon perhaps, but it has been used so l ons thut it y~uld be folly to attempt a chance. All vegetable cells with u few exceptions, have a surrounding wall of greater or leoa rigidity. The parts of the living cell are the nt1cleus, c~~toplasn, und chr Otl8tophoreo. Talrcn co llocti "le ly, these arc called protoplaom. The entire cell is filled with cytoplaom when young, but vacuoloo, or sapcavities form with age, gradually beco~ine larger and finally occupying the larger part of tho cell. The cytoplasn co~lects the nucleus and the cell wall.

Protoplaol!l \'Jes so nal!led by the Gernan botanist 'Ti/on !·.[ohl. It re- sembles the albun1en of an egg, being ol~my and viscid, and also sinilar in its chorniool proportioo. Protoplaom usually containo granules of starch, albumen, and so forth, which aro not conoidered eooontial constituents~

These grnnltloo l!lay bo soon eaoil~r vri th tho microocope.

It is more r, ropor to speal~ of pratoplasrn as a ~echanicnl r.iixtt1re chonioal compound. Ito chemical conpooition is very complox and varies oonotantly. Active protoplaso contains from 75 to 95% of wnter. 11

Proteid3 form the basis of protoplaom, so we find the chcnical elements

Oxygen, Hydrogen, Carbon, ~ itrogcn, Sulphur, end Phosphorus preoont. Other chenical clement a, of couroe, are enoontial to the no1,.mal growtl1 of green plants.

Tho physiological properties of protoplas~- those of all \ organisms - arc motility, nutrition, respiration, irritability, adnpta- bility, and roproduotion. A discussion of thooe proporties would consti- tute a discussion of Physiological Botany which is beyond the scope of this paper. From the physical conotitution of protoplasn, we would expect cells to be spherical in shnpe and no they would be if free and unconfined.

YOtlng cells aro most ~lw~"'O p·olygonal. The character of grovrth of the

surrounding tissue will dotornine tho shape of the coll ns it boco~os older.

CELL DIVISION.

It is bolio"1ad thnt each part of a cell in dori ved frorl the corresponding part of the gerrn coll. Coll division .is of two kinds - direct and indirect. Direct cell division is rathor rnre and will be only briefly noticed.

DIP.E CT DI 1T! S!ON.

This kind of di vision is l~nown as ami toois and oonotir1os as

fragmentation. At one ti~o, this mode of division wao thought to be the

common one, btlt it is now l:nown to bo very rare. In t;enernl, it mo.~r be

considered as talc ing place in old cells. It diff~ ers osnentially from

mitosis in that tho nucleuo re~aino in the. reoting state, no spircrno or

c hromoso~oo being formed. Amitoois in the higher plants ohould be oon-

siderad a purely socondnry proceos and not a ourvival oi a prirn~tivo process. MITOSIS.

Indirect cell division ia sornoti~os spokon of as karyokinesis,

though the later term, ~itosis, io tho one now generally used. This is the almost universal mode of coll division in the higher plants. It is also found in sorno of the lowor plants •

. ------STAGES OF MITOSIS.

For the sake of convenience, we divide the process of ~itonis into four stages, which, however, are not well defined. · The first

stage is tho prorhase; the socond, the ~etaph~se; the third, the anaphase; the fourth, the telpphaoo. The nucleus is generally considered the prime factor in most all vital prooessoo of the coll. In its resting state, or ordinary voge- tative condition, it usually appears as a rounded sec-like body. It is

surrounded by a distinct membrane, and c ontaina a conplicated netv1orlr of k,. lin'n throads among which nro nunerous crornatin-granules. I\

Prophase. Wban the cell preparos to divide, the nucleus undor goes both \, physical and chemical chanGeo. Tho cromatin aubstance looes ito not-like /\ arra.ngenont and gives rise to a thread known as the sp irc~e which breaks up into a definite number of pieceo known us chromoso~es. Each spocieo has a oharactoristio nunbor of ohromosomoo which is even in thono which tv multiply by sexual procesaos. 1Jt1ring the formation of the crornosomcs,

the nuceolus and tho nucleur wall disappear. The nuolenr ~e~brane," in the meantime, is surrounded by a fibrous layer. ~his layer is raised up fro~

the nuclour membrane ut tvro opposite points - the polar caps. The ont ire .. - - ~- ....

cell is now 1rnown as the mi to±ic fit;ure.

Metaphas~. The prophaac is really a preparatory process; the meta- phase being the initial phnao of nctual division. At this time, the chrosomos arrange thensolvea in a nore or less distinct plate which occupies tho equator of the dividing cell. Each chromoso~e now di- vides longitudinally into two exactly si~ilar halves, which finally go to oppoaito poles of tl~c SI>indle. The free :fi-bres run from polo to pole; those attached to tho chromoso!!les to one polo only. TJ10 splitting of the chromooo~oo and tho consoquent going over of exactly one-half of the chro~atin substance to each daughtor~nucloi is the most significant and tho esoontial fact of n itoaio.

AnaEhase. After tho chromoso~oo split, they begin to ~ovc towards tho opposite poles of the nuclear spindle. The spindlo fibres contract and thus draw the attaohod chro~ono~oo towa~do the poles. The fibros between the t\vO sets of ohromosonco aro l!:nown as rnantle fibres. Theso fibros show a series of deeply otuining thickenings in the equatorial plane which ultimately fore tho cell plate or mid-body.

Telophase. ~his, tho final phase, is not sharply scpareted fron the proceeding phase. Ao the chrom osomas approach tho poles of the spindle, they beco~e bunol1ed and finally grow togethor ond to end. A oint;lo filament is formod and this gro.dtially forns the restinG nuolous. ':1ho nucleolus and tho nuclear ne~brane are now formed and with the com- pletion of the oeparutinG wnll we have tw-0 duughtor-colls complete. T" =--

r.i.i to sis t aJ::os plaoo differently in so~e pnrtict1larn i n alriont ever;.I :p lant, so there are nany cxceptiono to the p~ocess ao outlined

above. To bogin to note exceptions would, there f' o~,.e, load to a papor of too great l ongt:-.. Onion root tips cut four and six ,f< thicl>::, and stained with Flecing'o triple stain were used hore ~ or observing t he process of vegetative rnitoois.

PLANT REPRODUCTION.

Under this head it is proposed to notice the gradually

incr eas1 ing c omplcxi ty of the plant body in the ascending scale of plant life and the accompanyine oherecter of reproduction.

The natural syste~ of classification of plants is based upon their actual relationship and whi1o it is not always p osa ible to determine the phylogenetic connoction,the system is none the lonn i mp or- tant. Even Linnaeus hin sclf felt thut such a syston was desirable, but the beliof in the irnnutability of opocios and tho very ~eager knowledge of the lower formsof plant life and the minute structure of the higher plants precluded . the poooibility of any better oynto~ than the artificial, soxual oysten propoeed by him in 1735. Linnaouo distin- guished twenty-four cluosos; t~onty-three classco of P hancrogu~ s. and one class of Crypto go~a. Wo still distinguish between Phancrogn~s , or flowering plants; and Cryptogams, or floworlens p l ants. On account of the comparatively oho~t tine ainco the natural classification of plants *11 15

has been seriously utte~ptcd and the oany uncertain points in connec- tion there~ith, it is only natural that few authorities agree on all points. ~ e r110.ps the wor1:: of, 'Engler anC. urantl io the best of i to kind.

In thio discussion, no uttcnpt ~ill be r.iadc to show tho connection between all tho variouo groupo of plants, but a typical plant of each

of the most i~portant c~oups will bo noticed.

The lotvost tmc1ot1btcd plant forms arc the Sohizophyta. TJ1ey possess powers of renintanco to l:ent und. other conditions ~1hich aro

fatal to ~ost o~ganis~o. ~his, together with the oxtre~e si~plicity of

cell str11cture, CllllSes us to believe t 1-.1 em to reprooont the

type of plan~s. Thero are two claoaos - the Sohizomyoeteo, or Bacteria, and the Sohizophy-oeae, or Bluo-Green Algae. Tho first class is destitute of chlorophyl while tne second comprises forms with chlorophyl. Bacteria are rniUM '. te 11nicell11lar plants - the sil!lplest of all

o~ganisbo. Tho colln nay bo iooluted or they nay f orn colonies.

The evidence now uva~lable is ngninot the existanco of an organized nuc-

leus. The method of reproduction ~o by fission or binary divioion, and by

.sporulat ion. The buctorilrrl ~irot incroasco slightly in oize, and in t hon

into tv10 nourly oqual po.1--ts by a gradual oonatrict ion in t110 mi d- o to tho ooocing procosn of higher plants is seen in

orlt.lnti on. Tho spore .¥.. arisc.1 w:.. thin the cell. It f irot appearo us a small listeninG spook gradually aboorbing into itoolf the protoplannic contents of the cell. Sporoa are highly reointnnt to hont, drying or other ad- Nothing that oven roaeoblea sexual reproduction is foo.nd

bacteria. Oscille.toria bel ongo to the rl°oc ond group of Schizophyta - the

chizophycouo, or Eluo-greon Al gae. It io found ovcrywhoro in water or dan p It is of tho simplest filanontouo fore. The filanento vi:ich bond

from· side to aido - ~1honoe tho name - are uov_ ally provi dod \ii tl1a

and cono!sto of oi~ilur, flnttc~ed cello. Tho f'ila~ent Depar~tcs into pieces, h oYmogon~; which grow into r.. o\·1 filarnonto. Tho center of tho cel l consists of a colorlooo central body ~hich correop onds to a nucleus and in the vc gtativc cello io in a otuto of continuouo division. Reproduction is exclusively vcs otutivo by coll divio: on. Rosting sporoo are formed by tl:e enlar g o ~ ont of oingle cells. reproduction of both Bacteria and ?lue- depondo upon fioo:on.

Tetran :e_ ~ra ~c p rcnento one of the oioploot for:r:is of tho Chloro yJ !l~r c-

Groen i\.lgao·. It io unicoll11lar, colonioo of \17hicn e ro ::nbodded in n gelat.nous matrix w~ich f ormo a flat groan thallus. nepDodntion io by ~cnno of ve g e~ntivo cell division - tho nµoleuo dividing by nitoois ao in t ho higher planto. 3ooidoo t~in opccial non-sexual reproductive bodioo, zooop oros are developed. It may ulso dc"T1clo:p simple se)~ual cello.

Spirogyra 10• ulno one of the Groon Algae. It io found va r y commonly

frosh ~ater. It conoiota of un unbranched greon filacont of olo~ga tcd cy- lindrical colla. Tho cell 0~ 1 0\10 u thin cytoplas~ic layer w~ich linoo the

contains ono or more flot opiral chromutophoreo in w ~ ich we find very plain pyronoids. A lurgo nucleus is found near the cnntor of the coll, stranda of protopluon run froo it to tho peripheral layer. Apparently

cells of the :!:)ilw:ient, lil:c tho3o of the plants alroud~T not iced, li "'le independently of one another. ~uch coll secures tto own food and each nuy

to forn daughtor-co~lo. Since it io difficult to sny how nuch, n cel l or u filament, formo un individual, we cnn not say uhothcr cell divisi on io growth or reproduction. If tho cell is the individual, its ui-

"lision r!lt1st bo considered :::-cprod~1 otion; if the :'ilnnento id the indi vi duo.l eel m... ~11 cre io. u d iotinctl:;' reproduct i ve procoon , h owe ver ,

tho cello t ul:o part. Th o collo of t\·7 0 ne u r :b~:r- filanonts nund ou t l a t. - +1 ., . h ...... h ...... + h era1 g rOYl \I . l S. Wn1C C OnJ 01.n v UO 11TI1 vl.ng "'· e cells in pairs. ~ho contents of one cell now puooea over into the other. The fuoion of tho ganctoo in

f ollowed by the f uoion of tho nuclei. Tho coll now be c o~os invested

a t hiclr \7all and in thio f 1 orm carries the plant over i1eriods of.. droug11t winter. It will be noted that the form of tho plant body - aonuming

filament to be the individual - is vary oinplo and that the ce l ls ~m ic h uhite in reproduction nre oimilar.

Vauohoria differs ~rorn Spirogyra in f orm in thut the fi larients

,...ur onc l"\.u o d • r+v 1s· occas::.onn· ,..... 1 y 0:~-'-+ uac · h o d t ·-o tho sub -s t rat urn b y co~or , i ess quite often· it io freo floating in tho water. Tho long fila- ants have no di7iding nulls, but are l ined with a protoplaornio layer v~ich contains numerous nuclei and chromatophores. Thero arc tno roothods of re-

Tho firnt is b ~ Deans of a zoospore wh i ch is formod in the ond

a side branch aftor this part io cut of f by a wall. Ono ~oosporo is f ormed

each ~oospora.~. iuun. The · ~cooopore is motile for a timo, then g orminate throwing out one or noro tuboo \Wiich develop directly into new plants. ds math od is lil:oly to prevail when the plants are grow·ing in water. Out f the water reproduction taJ:es place in this way: ahort, t hiol: branchoo rise on tho thallua. Thooe are ct1t off by sop ta and. become the oogonia.

of the oogoniu.m for~ tho egg-cell and an opening in the cell all appeurn. s· lender branches grow -out nco.r by and the e:Jrtrerni tios aro cut ff to for~ unthcrid1a.

,, Antherozoida, which rooonblo s~all zoosporoo, are finally oot Theoo mako their way - through a drop of dew perhapo - to the opon-

of the o~goninen, and one enters, fuses with tho ggg :~ -coll, and f orms

oospore which io finally set f roo and gorninates. It is thuo aeon hat a more complioutcd f aro o:, roprodt1cti on is ::~ ound in ~ro.uchoria. Chara belongo to a well defi ned group which shows a relationship

Siphoneao, of which 'l auchoria is o. reprcoontati ve, bl t they do to be oloooly relatod to any other group of plants. They nrc frcoh and may attain u height of more than a foot. They aro nt-

the sub-otrutUI:l by neano of branched rhi~oids vfr:ich opring frc~ at the base of tho a::t:os. Whorls of short branches aro g i ven o::f

short nodca whicifltcrnato with the long nodes. A lateral a.xis re- , the as.in axis is produced froc.. tho axil of the oldest s ide brunch of whorl. The aollo may be very largo in ~hich caoe the nucleus under-

repeated direct di v:..o :: on rooul ting in numerous lar~o nuclei. I n t l1e

a oinslo largo nJ clous, of the ordinary form, ~ich ividea by m ~ tosis when new cells are formed. Growth io from an apical

11 from whioh other cells . a~e ouccesaivoly cut off by the formation of ansverse walla. No special non-sexual spores are f ound. Bulbils may arise from nodes or roots and sivo rise to new plants if the conditions are right.

ia pr ovided for by the production of egg cal ls and ermatozoids. The anthoridin and oogOnia nre visible to the nalced eyo. Tho roplaoes a leaflet and the oogonium is the outgrowth of its sal node. The oogonia c ontain a largo .o7ul.e which is ourrounded by five irally wound tubular colla. ~heso oellc terminate in a crown through v~ ich ... e. sperrnatozoids enters. The v.r all of tho v.ntbermdiwtris formed by eight at cells from the lQlAter of each of which a large cylindrical coll nubri um1 projects into the oontnal cavity. The manubrium ar long septate fila~onto from each cell of which a opirully wound ailliato aporma1'ozoid is f ormod. The ooeporo,or fertilized ogg, on ger- riso to a simple filamontouo row of cells, tho ,- Rrom the first nodo of this ~izoids are produced, while one 19

or more main axes grow from tho socond nodo, and finally develop into n full grown plant. In one speoiea, the ovum can develop purthonogonet- ioally. It should bo noticod that the fertilized ogg coll becomes dotachod from the parent plant beforo germination takec place.

?darohantia ia one of the coornonest .Li vorworts. Unlilce the Algae already noticed, it io a land plant; preforing damp oituationo, ,-z:.. however. It hao no true ruuto, but in supplied with rhtoids, or absorp- ~ tive hairs. The alternation of goncrations characteristic of\ thio am higher olasseo of plants is plain in the case of Marohantia. A 3oxual generation, provided with sexual organa, is knewn ao tre G~~etop~ytc; an asexual generation, producing spores; .is. lrn.own ·as. tho s~orophyto. The sexual generation arisoo froc the sporo; .the asen10.l, from tho for- tilized egg. The number of chromoaumeo wn the nuolei of the sporophyte is twice as groat aa in the nuclei of the ganetophyte. The gacetophyte io the predominant phaoo of llarohnntia. It io a forked thalluo, the branches of which grow forward while thooldor pa.rt dies, t huo separating tbe

and giving rise to now individuals. Pores on the OUIElit, a:i.milar to stomates, lead into ohamboro from tho :.. fldO.is ~~ of .'"- whioh rows of collo form the ohief Asoiumilstory tinsue, arioe.

Reproduction is of t-r10 1:inds. Gamrnae arinc 11pon the upper sur- face of the thallus, aro oct froo and develop directly into noTI plunto. _, Th is is vogetati vo propagation, and answers t!1c s amo purpo30 ao zoosporoo in t ho A1 gao, nane~y,, rapi·a nu1t1 p.1ca~1on. , · ~· Anthoridial and archcgonial bi.. anohcs· arise from the thallus. The anthcridiul brnnchon torrninutc in lobod dioco which boar the an- theridia on their uppor oidoo in flo.slr: s h aped dopressiona, oach contfil n- ing an antheridiurn. ~ho lars c cell of tho antheridiUI!l formo a nunbcr of sm~ller colls, in onch of which a single anthorozoid is : ornod. Tho .arm- egonia arc found - ~ n rowo botwoon tho rays. Tho ogb coll of the archo- gonium is fertilized by an unthcrozoid and tho resulting oooporo di- vides repeatedly, tho na0 0 of co l lo thuo originating finally f orming a stal~ed oval sporog oniun • . numerous, freo, somewhat rounded oporeo together with a largo number of fino thrcado arc sot free from tho apDrogonuim. The germinating aporoo rise directly to tho thalluo of tr£ new plant.

l:llnium io tal~on ao n reprooo!! tati T.;e mass. The otor.i und loaves a 1"0 differentiated. ~ho otem io ~ornod of cells which gradually bocone s~allcr and thicker wul l od towa r ds tho periphery. A central axial strand of olon- gated, conducting co l lo with narrow 1Ql:lina is found in tho 0tom. Those strands, h owever, aro not ao h i ghly differentiated ao the vaoculnr ~un d lca

.of ferns. Tho lea ~lcs ha.. 10 a "'Jory s ioplc structure - eonsioti. nt;

• of a single layer of cells. The rhi~oids aro similar to those of I.larchantis.

Reproduction io coocntia ~. ly the saoe as in I~archuntio.. ':ho sexual organs are terminal on tho ~ain ro~io. The ~ntherozoid io motile by nouns of t wo cilia, roachinG the OGg-coll uhon t ho planto aro wot. The oporogoniun is a rathor complicated structure. Tho germinating np oro dooo not s ivc rine directly to the J..'.r oos shoot, but to a nuch branohod fila.nontouo gr~'th l:noir1n as t he pi:_otonema. ~ho ~; ounG nooo ohoot io developed fron omnll budo zfuich arise fron tho basal co l l of one of ito branchoa.

Aspidilltl?l is a truo forn - a representative of the most hi ghly de- veloped Cryptoga.nes. IIore as in t110 lo.ot two p lanto noticod q dintinct , alternation of s oncr ationo io s.--iov1n. Tho soxual generation bears anth er- idia and archogoniu; tho aoexual 'i:ich developoo froD the fertilized egg , produces asexual, unicollula~ spores. The sexual generati on uriees on t~ ger mination of tho apore. ~he sporophyte is the conopicuouo phaoo of f erna. Tfle s~me to ph oro is u nmull, thin, hcart-ohupod body culled tho p rotha l ~iurn. fJ2 1

It is attuohod to t110 ground by r11izo:.ds and . roocribloo a onall ~hullcid l i ver\vort. T110 anthorid.ia aro borne on tho lov10~ ourfaco near tho more pointed end of the protho.llit10, ~h:.lo tho archogonio. arc fo11nd nco.1~ tho notched oxtromit1. Archcgonia and antheridiu arc borne on different pro- thallia in oomo opcoien. Fortilizution tal:eo place v111 en tbe prot110.ll ium is v1ot vii th dow or rn:.n b:>· the entrance of an anthorozoid :.nto tl:o urc11oc o9'."' nium and the conjugation o: tho egg-coll and the anthoro~oid. After for ti- lizat:on tho es s-oell dcvolop3 into u multicellular enbryo vhich bcconoo the sporophyto. The oporop~yto io a plant ponnossing ~ highly differentiated internal otructure and externally scg~onted into 3ten leavco and true roots.

The fertilized ogg-ocll dividos repoutodly and forms - nithin tho arche ~ on ium - the stem apex, tho firat loni , the pricury root, ~nd an organ ~culiar to the ? tor i d ophyto s, tho oo culled foot. Tho ;,r oung 3porophytc dori vo a it o nouris111lont fr on tho prothu1. lilt·:tn by r:ieans of tho foot ti 11 itn onn roots are able t .o 0111;p ort it. 'l1hc otomo loaves and roots of the sporop11yte a r e traversed by well di~forontiatcd vaocular bundloa. Sporoo arc borne vogo - tativoly in oporang ia ~h~ch uro cluotered on t~c under a~don of t re leave s. Each ol uotcr, or oo r~ o, io ~~iolded b y a oenbranc tho induo~ ur1 . The sporang ium ccnoists of u wull enclooing tho opor~gonouo tioouo, tho opore mother oells of uhic~ give riao ouch to four totruhedrnl op oron. Co~par cd with r!archant io. o.nd ~1ni un tho 3porop h~rto of 1\opidiun c orro n11 ondn to tr.. o sp orog onium. Tl10 prothallitun of tho fern en tho other hand corrcopondn to t ho protonoma and o11o ot of :.1nium.

zea lil:o moot I.I on o cot~~lodono iG herbacoouo. '2:' I'-c embryo 110.n on l )l one · cotyledon, und tho ultornnto loavco are arranged in tTio ran1~o. ~he roots are fibrous. ':1horo io no c lcar d:otincti on of pi th, ba r 1:: and '\1 ood, co ncontri -

. + ~ ~n n cally a rranged. rho oporo p h~I to io t ho prodoninanv p .,..aoe, - f-U.nct, u. casual obser vation would not reveal tho ganctophyto. Upon gorninntion tho rud:clo and hypocotyl of tho snull embryo omeTge from tho seed cont \Veile tho co- tyledon remains within tho oecd. The growth of the root io urrcotod and ito place ta1~ on bd.. nurnorouo ad··v·entitiouo· rooto \7hich opring fron tho s tern v1b.ich is unbranched. · The bull~ of\ it

the axilo o::· tho loavoa. ~ '1:ho gurnotophyto is veI·y incons1):. c11ous and ~uy

said to be parasitic 11p on. tho op oroppyte, the eribryo of tho ne\1 oporor)l1;/ t e developing bo f oro tho oood io ~reed f ron tho old. Since tho gonotophytc docs not difi1 cr matorio.llil from tha t of Co.paella \711i ch is to bo doocribod in some detail, it ,,,,ill not bo ft1rthcr r 1 oticod.

Capsollu is tal:on no u ropreoentati vo Dicotyledon l'::1 ich is t11c pro-

plant type of today. ~ho '. nltornation cf goncrntionn notiood in

Marchantia. and .&t np idiurn is prooont hero t 11 oug1'1 it i s t110 nporoph:lto -r,t. i ch the predominant phaoo; in fact tho ganetophytc i n vory in4onspicuoun , a nd

seldom t h inlt: of i t in connect:. on

ste1n and. loU'"'Joo. r:''hc prir:iary root peroists and throv1s out numer·ouo

branches which b oar ~ root hairs through which water and f ood oatorials

• are t a1~en in. T·ho br·anchod otom :.3 di fforontiatod into pith, wo od and / r:i ho pi th f oros the cont er and the ba:rl:: the outer layer, the wood

\ bct\voen the t\10. Gro,vth ta1:es place in the car:ibiun layor junt t1ndor The lea7oo uro nett .ed veined, and the nnrbi ns are cut. Tho hermaphnodi to floworo o.r·o bor-nc in u long racer.no. Tho dovo1Qpnent of the flower, which .is the noot chnruotoristic

of all 11-n g :i.oopcr n n, '7ill ba :~ broifly noticed. Tl1e t ypical fl o'vor consists of a serios of\ f lor al leaves arranged nb ol1t tho apex of the s h oot or floral axis. :ho outornoot loovoo, or SeEals, aro protective in func- tion and together oonotitl1to tho Cal~~. Tho .second oories, tho .::·otnls, col- • lect ively f orm tho Corolla ~r.icn io usually conspicuously colored and servos

. 'I.. to rettract tho attontion of inoects Tihich often effoct pollination. Tho st u- . mens on which arc borne t ho pOllon oacks come within tho corolla. Collect-

tho Gt amens forn tho ~~ ndroocium. The innerrnoot sporophy~ls, the carpels

separate, out as a rulo, they farm a compound piotil. Tho car1J ols t al~ - together form tho Gynaociun. The rnicrooporangia, or pollen oaco, make

anthers of the otan~. ~ho ~ncrooporang iW!l or ovule i s borno with- ovary which is tho basal portion of the piotil.

Tho flo~al loavos arc iroducod on tho roceptncle juat an foliage arioo from tho growing point of a vegotativo shoot. Tho sepals

first, foll ouod by the otamens, carpolo and potals in order. 1~o

j rowth in length of the floral axis is arrosted by the formation of the

The :macrooporant;iurn, or ovt1lo is an outgrowth of t110 carpel. T he

from which i t g rO\'JO io 1:noi;1n as the placenta. The rnacrooporungium onsists of the nu~ao, f:rom tho bnso of which integuments ar i se nncl cn~ar g e ill they lcavo only n onall paooago, the micropyle, which loado to the upper

. I r micropylar ond of tho onbryo-oac ~ tho opposite ond is called tho nnti-

ond. Tho ct1bryo-oo.c arisod from a single hypoder:r.ial ce 11. From t 11i s an outor cell, tho t~otuo is cut off. The inner cell divides oovcra.l imes, one of tho cello grows faster than the ·othors and dostroying thorn bc- omes t he oobryo-oac or nucroo:pore, uhich is at first completely :filled m t h rotoplosrn containing a coll nuclouo. The two caturation or reduction di-

£;Jnd tho noxt di ""lioion all of' which t a 1ro place \7 i thin tho cr.ibryo-snc to tho f ormation of the egg rnuot now bo noticed. Tho reduction divioi on

leado to the f orr.io.t i on o ~ the f o11r primary cello of the onbryo-oac is /fa 24

OOf£eno nis, the rodt1ct :.on di vioi on v:hich proceeds tl1e f 1 ormuti on of t he pollen grnino is called opormatogonosis.

The oingle nucleus ~f tho arrtbryo-sao we will asouoe to bo in tho resting otago an deocribod ~oforo in the case of vegetative cello. The Y::i~) chrosomcs bocon1e uni tod in pairs in tho prophaoe and this is foll-O\"lod b;I a ,ft.. marked ccntra.ction of tho nuclear contents which is characteristic of this process of division and is called synapoio~ollowing synapois the double chromosomes again aopurato out uo a delicate double filament. This soon unites forming a stout thread which io arranged as a loooo, skoin. Again

the doublo nature of thin t11reod cnn be nado ot1t ..: ... ,, ._ ·~ : and it now falln into segments, each segment corrosponding to ono paired chromooome. Since two ohromosomco are represontod by each segment, tho numbor of aogrncnts is only half that of the chrornosonos of tho t issuo cello of the plant. Tho JR ir·od chromosomes ha ving become ohorto:r and thicl ~ er are d.istributod arol1nd t11e of the nucleus; this hao boon called diakinoois. Einoplasmic

aro now applied ~o tho nuclear membrane which n~; disappears . • !he nuclear spindlo - at first mul tipolar, ultimately bipola.r - or~f;fn&tea from tho lrinoplasnic fibreo. The !fibres of tho spindle become attached to ohromoaomoo now arranged in a equitorial nuclear plute. Tho of tho chromosomes, which were before united in pairs, soon place. Hare tho oooontiql of the reduction divioion io effected, for

not longj._1udinal halveo of chrornoaomos but entire chronosonoo v~ich

epar.ate from one another. Eaoh do.tighter nucle110, thcre:f ore, roco :!. .,.;en only

many chromooor:ien an o.ro f otmd in tho tiooue cello o:f tho, came go A longitudinal split can be oeon in each chrornooooe as they/toward poles. Tho formution of tho daughter nuclei precocds ao bo f ore dooUJribed

• vegetative cells. Thin di 'Tli. oion knov1n us tl10 heterot;y1?o di ·vision l.S tOm}?ltly folloucd by a oecond, or homntype, divisioh. In tho hornotype divisi on

o nevi long i ttldinal spli tt:.ng 0 ±1 t110 c hrornooorie o tal::e a place, but the tv10 alves of each chromosome, which oxiatod in the daughter nuclei become sepa- 4 from one ·another, e..nd bocone t110 chromosomes of tho grand-duughtor. nuc-

The hornotype di vision ogre co in other respects with ordinar;{ nuclear ivision. One of the charactoriotic features of tho wt-olo procoos of matur- tion or reduction divio i on io tho rapidity with which tho divioion.Ssuccoed another.

Thio t a "brief deacrip: ion of oogcnosis t bringo llD to tho four

cells of tho onbryo-oao. Two of theso oolls aro found nonr tho oi- ropylar ond of tho o!!lbryo-oac and two near the antipodal e·nd. Each c i 1 the rimary cells nou divides - one at the n ~ oropylar ond to f orm the two syn- r gidae; the other to form tho ogg und the upper polar coll; one at the as-

coll; one at tho antipodal end to t\vO of the three antipodal e·ollo, the other to form the third antipodal and the louor polur coll. The upper and loTier antipodal cello conjugate

o form tho ooc onda.ry or.ibryo-oac-nt1cleuo, by the di vi oion of v.?hicl1 tho ndosperm .colls arioe. Tho throe antipidal cells develop cellulose wollo part in tho ±'t1rthor dovolo1;ment oil tho oacroo1;oro, finall y do-

Tho oyncrgidno which becone i nvestod with thin cytopluonic rne ~b- dcgonurate eftor fertilization of tho egg. Of the oibht cello ising as described, only tho ogg contributos to the rnorpholoGical formation embryo. The nole ooxual gencrot:on will now bo briefly noticed. Tho mi- rospores, or p ollen-grains, uro f ormed in the mi crosporang in or pollen-sacs • hich develop within· tho anthoro. Tho pollon ones are dovolopod : rom the ypodermal layer oi' cello. The a :r- chosporium under goco a nlmbor o:f divisions

nd forms tho p ollen ~other collo. Each pollen mother coll now divides tvace

arming four poll en g rains. Thcoo arc the oo.tt1ration di·vioionn nnd ure own as opcrmutogcnoois. Si11ce they do not dif'fo r i' ron oogcnoois., an a 1J o,1e ascribed, t ho; will not be described. Tho pollen gr uina arc spherical in

hape and n~o provided with a cell wall. The wall conoiots of an external

ayer, the exine, and an ~nnor l uyor, tho intine. Just uo tho f our primar y

el-=. s of tho ombry o-oac di vido f t1rtl: cr, so t h ere are two a tl .-: ~ ti onal di ·visions

tl1e pollen gr uino. ~his will bo noticed t1nder p oll ination, a dioc11Gnion

v1hich :fb llOV/ O:

Tho p ollen grain3 arc carried to the otigmu of the p isti l by ~and

insectG ~ n most caoea. A socrotion is gonorally developed from the tigmatic ourfaco nnd th!o otimulatoo the germination of the pollen gr ain.

e germinating pollen g ru~n io oeen to consist of a vo~etutivo coll anu

antherid~al mother-coll. The vegetative coll growo out uo t ho pollen ube t he wall of which is continuouo with tho intino. Tho pollen tubo rupturoE

he oxino and p onet~ntco :!.nto tho tioouo of tho mucrooporangiun und grown r ap- idly through tho style, t ho col2.o of v1hi c l1 no do11bt, furnioh t re natcr ial

for its growth. ~he pollen tube advanceo until it roachoo tho icropyle of tho ovule, into Tihich it ponotrntos, and puoheo through tre

issue at the apex of tho nuoolluo nnd ontors tho embryo-sac. I n t he aantin10, tho antl1eridial nothor-coll, which has pasood :!.nto tho p o2.len tubo,

tY10 gcncrati 'Tio nt1cloi. Tho p oll en tube us11ally g1-- ows

hro1igh ono of tho o~.,.nort;:!.d . o.o, ·. whicl1 is dootroyod, and d:i.ochargco one of generutivo nuclei into t ho egG . whore it f uoos Ti ith tho egg nuolcuo.

second generative nuclouo ~o diocharged into tho enbryo-onc and ~ay .. tho enuoopc rm nuclcuo.

The f ertilized C8G f orno a second cell, t h an othe~n ~ n a linour or in u more conplex chain, known as tho proonbryo or ouoponnor,

ter1ninal coll of \"/bich the initial cell or cello o: tho ombryo

Boaideo tho ohangco deoc ~ ibod in tho cnbry o-oac oth0ro are g oing

in the nucellus and ito intcb,unonts. ood is otorod in tho one or -:ho dc·voloping tiob110 a1~011nd it , no.1:in[; 11p

he al·bumcn o f'i t110 oood. Tl10 changes g o beyond t110 o'T111 :!.o n r; i t 1.. ipeno into

seed, and thoovary , and in oono ~notun ocs aooociatod partn , bocono t he •

Tho mu cr~op o~o o ~ the hotorooporous ~ toridop hyton in ro p~ cnonted

the ombry o-oac, tho ganotophyto by tho otructures dovelopou within i t . tho above case of Cupsella, a roprosontativc Dicotyledon, the asexual . eneration .-u- ("! vo-::y oomp lo~~ in forr:i and higl1ly organized while the oox110.l eneration 10• reduced. T~e aooxual plant encloooa the female sexual soneration throughout its entire dovolopncnt which only be- omes separatod froo it in tho ocod which contuino the co~nenconont of

succoodins uaoxual gcnora ~ ion uo the embryo.

Tho a ccompany~ng diabruno arc designod to ohow the gcnooin of egG and tho scncrutivo-nucloi. m Primary mother pollen cell. C Secondary nether pollen cello. O P·ollon grains. Et Ve getative cells. <:) Autheridial mother cell o. • Genorati ve-nucloi.

I ~ iiother coll of ornbr~lo-sac.

Uppe·r DaughdJor coll of OI!lbr.fO-SflO. EB ' "" ,, Lovier . " " D· ~ " " O Uppe r piimary cello. " " " e Low-1er " " " "

e o~Tlll!l or egg @Upper polar nt1clous OLo\ver " " S)Synort;iduo 0 t ntipodal cello. PlillT III.

S~oto~atio Botany.

The v1orl:: in Syotonutic Botany wao bogt1n !.larch 1, 1909 nnd con- tinued tilli--Juno 1, 1911. Tho appended list o:r1 ows 41'7 speciniens col1e ctod, end identif1 icd, ~!)of v1 h. ich were not mounted. Thone not ncuntcd are r.iar1:od

Vii th a * • About half of thia nunbor woro injurod by inoccts or nould to such an oxtont thut thoy '\7oro uni1 i t f 1 or m·uunt ing. The mould could h u\e b een prc""\t7 cntod, or c our so, bt1t during tho sc_r.anor, -r1i th onljtT nev1opup cr a f 01-- driers, and ao many an t-r1ont;/ opocir:1 eno put :!..n the i)roos at one tine, oo Tje of the opooir.iens ".1oro ruined bof~oro the inju_ry \7ao detected. Spec in1ons in- jured or dcstroyec wore replaced whcnovor pooo~blo, but thin coul~ not al- ways be dono bocuuoo of tho habitat was too far removed, or tho fro\1oring season pa:Jt. Tho rnil1:":Tceds, gentians, and sor10 conpooi tao \'1erc badl~r at- tacJ.:ed by inne cts, and t hio a cc ounto for some miooing opocineno.• Done. uro mi ssing on account of mounting then - the conifcro uro among these. Again, others ";.~ ere not I:lOt1ntod beca11so of tho very v1oal tb of material, jt1st ao soIJe plants, po.rt :i.c11lnrl~Y'· culti vo.tcd oneo, wore not e·vo n worked out. As a nuttar o: fact, very few cultivated planto Tioro coll ected, except tho grasoco and f ru:ts. ~he cultivated truito uro includod becuuno

I of other work done with thon. ., Gray' o "11ev1 rJanual of Botany" and Britton and Brown' o "Illt1str a tcc

of tho Northern States and Canada" have boon the principal ~ anuals used, the for~or hao boon -foll owed in the classification and nornonclnturc in those caaes whero tho two did not agree.

]Jore than half , 223, of the specir1ens ca.me froo :"~ t1l u slti Cotlnty.

Ii!l ontg o?!lery Count;; contributed 1?7 ,· and Viythe County 17. ' The uppendod lint nha; o 87 fa~ilio s und 273 genera roprooent od . i ncluded 7C Conpooitne, 34 Labiata o,

24 Logurn:iosao

22 J ooacoao 19 Scrophulurinco uc 13 Cruciforno Raunnculncoao

11 C ru~inoao, Lilincouo 10 Ericacoao, 1iolacouo 9 :? olygonucouo

8 l slopiadacoaorn ~ary ophylllu~eae 6 Borabinucno, Orchiducao, Solanacao

5 Lobeliaceao, ~ rimulaceae, U~belliferae

4 ·3erb.aridaooae, @apifoliaoeae, Ettphorobiaceae, I'.ial vaceae,

onagraoeae, ~) lunta c; inaccae, Robiacoae, Sa.xifrar;nceao.

-~ '""' 3 Aceraceae, Cnnpaunlaceae , Chenopodiaceae, Gentinnaceae , . in~c. ea c: , Polyr;alaceao.

2 J1 canthacoae , f.1.I:lerantr1aceo.e, liristolochiaoene, T3a l sanmn c ea~, Betoulaoeae, 5ignoniacoae, Convolonlacene, Coruacoac, Cr aonula-

ceae , Fagaceae, Flm.arinccae, !~ ama.melidaceo.e, !iypcricaceno ,

Oxuli duceac, Polypcdiucoac, ~o rt u lacaeao, Filinceae, Vrti caccae ,

Vorbenacoo.c.

1 Alisnaccao, ~nury llidacouc, ~nacordi acouo , Ap ocynnccao , ~ ra c cao , Aruliaccac, Cclaotrnccoo , Cyp oracoue, Diapcnniucoac, Dioocorcnccae , Dipoacuccao, Eanoetncouo, Gcrumiucoao, r ydrophyllacoao, lllccobrncc[ I ridadeue, Juglonducouo, Lauraceao, Loguniucono, 1ytbnacouo ,

I ~ nguoliucouo, Cloucoac, Ophioglosoa eoae, Orobauohacono, ~ apavora - cone, i'h1ymacooo, ~hytolnccacono, r lntnneocao, :1 olon oninccno, 0

Rh nnnacono, Snpindacono , St aphyleaceao, St yrncaccno , TYJ.)hacouo .

• Tho diotribution of ~ ho opcc~ea and genera 10 nhov1n b ~l tho follorning table:

• 0 • • F o.1~ i l i c o : Gonorn • uPOClCS • • • • t Ptcriodph;lta: 3 '• 3 '• 4 • • '• •' '• '• '• '• '• t '• • • • Gynnos per- '• '• •' '• ' '• ~ac • 1 3l. , •' 3 •' '• '• '• '• '• • • '• ' :Ui ono co tylo - '• • ' •' • '• 1 () '• •' '• dons ' 30 35 '• • ' • '• t • '• • •' t Diocotylcdon:;;' 73 • 237 '• 3~5 '• • •' '• '• '• '• •' • '• •' ')'i 3 •' '• Total 87 "'-' . 417 •' '• '• '• ' Abutilon Theophrasti Medic. Ambrosia artemisiif olia L. :Ambrosia trifida L. Acalypha virginica L. Ambrosia trifida L. var.integrifolia (Muhl.) T.& G.

Acer Negundo L. Amelanohier canaaens i s (L.) Medic.

Ao er rub rum L. Aoer aacoharum Marsh.* Amphiearpa monoica {L.) Ell.

.. Acerates floridana (Lam.) Hitohc. Andromeda floribunda Pursh.* Aoeratea viridiflora Ell.

Anemone trif olia L. Achillea Millef olium L. Anemone virginiana L.

Aotillomeris alternifolia (L.) DC. Anemonella thalictroides (L.)- Spach.

Aesculua Hippocastanum L.* Anthemis Cotula L.

Agastache scrophulariaefolia Anthirrhinum majus L. { ~Villd.) Ktze.

Agrimonia s triata Michx. Anychia polygonoides Raf.

Agrostemma Gi thago L. Apocynum cannabinum L.

Allium cernuum Roth. Aquilegia canadensia L.*

paniculatus L.* Arabis canadensis L. Amaranthus retroflexus L. Aralia racemosa L. , Arotium minus Bernh. Baptisia tinctoria (L.) R.Br.

Arisaema triphyllum (L.) Schott. Benzoin a.est·iva.le (L.) ~Tees.

Aruncus sylvester Xosteletzaky. Barberis aanadensis Mill. Berberis Fortunei Lindl. Asarum aanadenae L. Asarum virginicum L. Betula nigra L.

Aaclepiae incarnata L.* Bidens bipinnata L. Asclepias phytolaoooides Pursh. Bidens cernua L. Asclepias quadrif olia Jacq. Bidens oonnata Muhl.

L Asclepias syriaoa L. Bidens frondosa L. Asclepias tuberosa L. i Asolepias verticillata L. Botryahium virginianum (L.) sw.

Asparagus officinalis L. Brassica campestris L. * Brassica Napue L. * Aster lateriflorus (L.) Bri·tton. Brassica oleracea :L. Aster Lowrieanus Porter. Brassiaa Rapa L. * Aster multiflorus Ait. Aster oblongif olius Nutt. Bromus seaalinus L •._ . Aster patens Ait. var.phlogifolius Nees. Aster prenanthoides Muhl.!

Aster Sohreberi Nees. Cacalia atripliaif olia L. As ter undulatus L.

Oaltha palustris L. Astragalus canadensis L.

Campanula americana L. 33.

Campanula divarioata Michx. Chrysogonum virginianum L.

Camptosorus rhizophyllus (L.)Link. Chrysopsis mariana (L.} Nutt.

Capsella Bursa~pastoris (L.) - Cichorium Intybua L. Medic.

Carpinus caroliniana Walt. Cimiaifuga racemosa (L.) Nutt.

Cassia marilandica L. Circaea lutetiana L. Cassia niotitans L.

Cirsium lanoeolatum (L.) Hill. Castanea dentata {11arsh.) Borkh.

Castanea pumila (L.) Mill. Claytonia virginica L.*

Catalpha bignonioides Walt. Clematis ochroleuca Ait. Clematis Viorna L. Ceanothus americanus L.

Celtis occidentalie L.*

Conopholis americana (L.f.)Wallro Cercis canadensis L.

Corallorrhiza maaulata Raf. Chenopodium ambrosioides L. var.anthelminticum (L.} Gray. Chenopodium Boscianum Moq. Coreopsis major Walt.* Chenopodium murale L. Cornus Amomum Mill. Cornus florida L.~ Chimapila maaulata (L.) Pursh.

Corydalia micrantha (Engelm.) ~ Chrysanthemum Leucanthemum L. Gray.

.. ...~ : ( t: .. : ~ • \. II -· ~ ...... - ~· • ; Cryptotaenia canadenais (L.) DC. nesmodium Dillenii Darl. De smodium grandiflorum (Walt.) DC. Cunilia origanoides CL.) Britton. Desmoaium nudiflorum (L.) DC. Desmodium rigidum (Ell.) DC.

Cuphea petiolata (L.) KOehne. Desmodium rotundifolium ~ (Michx.) DC ~ *

Cus cuta Gronovii illd. Dianthera americana L.

Cynoglos sum officinale L. Dianthus Armeria L. Cynogloss um virginianum L.* Dioentra Cucullaria (L.) Ber nh.• Cyperus ovularis (Michx.) Torr. Digitalis purpurea L.* Cypripedium aaaule Ait. Diodi a teres Walt.

Da ctylis glomerata L. Dioscorea villos a L.

Datura Stramonium L. Dipsacus sy1vestris Huds .

Da tura Ta tula L. * Diaporum lanuginosum (Miohx.) - Nichols. Daucus Carota L. Dodecatheon Meadia L. Delphinium exalta tum Ait. Draba ramosissima Desv. Dentaria l a oinia ta Muhl.

Desmodium braoteosum (Mi chx.) DC. Echinochloa crusgalli (L.) Beauv. var.longifolium (T.&G.)Robinson.

Echium VUlg,;,re L. Eleusine indica Gaertn. ~ ' Fagopyrum esculentum Moench.

Epigaea repens L. Fragaria virginiana Duchesne.*

Epilobium coloratum Muhl. Yraxinus americana L.

Epipaotis pubescens( ild.)A.A.Eaton. Galax aphylla L. Equisetum arvense L. Galium Aparine L. Ereahtitis hieraoifolia (L.} Raf. Gaultheria procumbens L. Erigeron pulchellus Michx. Erigeron ramosua (Walt.) B.S.P. Gaura biennis L. I

Erythonium americanum Ker.* Gentiana Andrewaii Griseb. Gentian& quinquefolia L.* Eupatorium ooeleatinum L. Eupatorium perf oliatum L. Geranium maculatum L. Eupatorium purpureum L. Eupatorium urticaefolium Reichard. Gerardia flava L.* Gerardia linif olia Nutt. Euphorbia oommutata Engelm. Gerardia pedicularia L. var.ambigens Fernald. Euphorbia corollata L.

Euphorbia Preslii Guss. Geum canadenee. Jaaq • Geun flavum (Porter) Bicknello Evony.mus atropurpurens Jacq. Gillenia trifoliata (L.) Moench.

Gnaphalium poyycephalum Michx. Halesia tetraptera L. Hypericum gentianoides (L.) B.S.P. Hypericum punctatum Lamo Hamamelis virginiana L. Hypoxis hirsuta (L.) Coville. Hedeoma pulegioides (L.) Pers • .

Hystri~ patula Moench. Helenium autunmale L.

Helianthus tuberosus L. Impatiens biflora Walt. Impatiens pallida Nutt.* Heliopsis helianthoides (L.) Sweet. Ipomaea pandurata (L.) G.F.W.Mey. Hepatica triloba Chaix• • Iaanthus brachiantus {L.) B.S.P. Hibiscus Trionum L.

Hieracium Gronovii L. Jeffersonia diphylla fL.) Pers. Hieraoium paniaulatum L.

Hieracium pra~ense Tausoho Juglans nigra L.

Hieraoium scabrum Michx. Hieraaium venosum L. Juniperus virginiana L.*

Houstonia caerulea L. Hou8tonia oiliolata Torr. Kuhnia eupetorioides L.

Hybanthus ooncolor (Forster) ~ Spreng. Lactuca oanadensis L.

Hyd~ophyllum virginianum L~ Lactuoa hirsuta Muhl. Lactuca sagittifolia Ell. . Lactuc~ villosa Jaoq. ··t.yc9pus: americanus Muhl.

Lamium ample:xicaule L. Lysimachia quadrifolia L.

Lappula virginiana (L.) Greene.

Malva rotundifolia L. Leonurus Cardiaoa L. Karrub:tum vulgare L. Lepidium campestre (L.) R.Br. . Lepidium virginicum L. Medicago sativa L.

Lespedeza hirta fL.) Hornem. Melilotus alba Dear•

Linaria vulgaris Hillo Melissa officinalis L.

Liquidambar Styraciflua L. Mentha arvensis L. var.canadensis (L.) Briquet. Mentha piperita L. Liriodendron Tulipifera L. Mentha spicata L.

Lithoapermum canescens (Michx.)~ Microstylis unifolia (Michxo)~ Lehm. B.S.P.

Lobeli& cardinalis L. Mimulus ringena L. Lobelia inflata L. Lobelia leptostaohys A.DC. Mitella diphylla L. Lobelia spioata Lam. Lobelia syphilitic& L. Monaraa diayma L.

t Monarda fistti.losa L. Lonicera japoniaa Thunb. Monarda puno·ta ta L. Monotropa uniflora L. Pedicularis lanceolata Michx.

Panthorum sedoides L. Nepeta Cataria L. Nepeta hederaoea {L.) Trevis,an. Pentstemon laevigatus Ait. Pentstemon laevigatus Aito var.digitalis (Sweet) gray.

Oake·sia sess ilifolia (L.) ats. Phleum pratenae L.

Oenothera biennis L. Phlox ova t a L •

. Orohie spectabilis L. Phryma Leptostachya L.

Orobanohe uniflora L.* Phys~lis heterophylla Nees. ' Physalis pr uinosa L. O:xalis stricta L. Oxalie violacea L. Physos tegia. virginiana (L.) Benth.

Oxydendrum arb oreum {L. ) DC. Phytolacca deoandra L •

. Oxypolis rigidior (L.) Coult.&- Pilea pumila (L.) Gray. . Rose.

Pinus 8trobus L.* Panicum capillare L. Panicum clandes tinum L. Plantago aristata Kichx. Plantago lanceolata L.

..,. Paulownia tomentosa ( Thunb. )~*'·:~ . ~~ -: ~ ii1antago major L. ,... Steud. Plantago virginica L. Platanus occidentalis L. Portulaoa olerac·ea L.

Poa compressa L. Potentilla oanadensis L. Paa pratensis L. Potentilla monspeliensis L.

Podophyllum peltatum L.* Prenanthes altissima L. eerpentaria Purah. Pogonia verticillata fWilld.) Nutt. Prunella vulgaris L. Polygala pauciflora Willd. Polygala Senega L • Prunus amerioana Marsh.* . Polygala verticillata L. Prunus avium L.* var.ambigua (Nutt.) oo,d. . Prunus Cerasus L.* Polygonatum biflorum {Walt.) Ell. Prunus hortulanQ Bailey.* Polygonatum commutatum {Ro& s.)~ Prunus serotina Ehrh. Dietr.

Polygonum aviculare L. Pycnanthemum flexuoeum (Walt.)- B.S.P. Polygonum ereotum L. Pycnanthemnm pycnanthemoides- (Leavenw.) Fernald. Polygonum Hydropiper L. Pycnanthemum virticillatum- (Michx.) Pers. Polygonum soandens L. Pologonum tenue Michxo Pyrola rotundiflora L.(B.& B.) Polygonum virginianum L. Pyrus aommunis L. * Polymnia canadensis L. Pyrus Cydonia L.* Polymnia uvedalia L. Pyrua japonica Thunb. Pyrus Malus L.

Polystichum aarostichoi~e· s ::_ ·· - .~ ~~ (Michx., Schott.* Radioula Naeturtium~aquatioum~ Rumex odtusif olius L. (L.) Britten & Rendle.

Ranunculus abortivus L. Ranunculus hispidus Michx.* Sabatia angularis (L. ) Purah.

Raphanus sativus L. Sagittaria longi•ostra - (M.Micheli) J.G.Smith.

Rhododendron maximum L. Salvia lyrata L. ~ Rhododendron nudiflorum {L.) Torr. Salvia splendens Sellow. Salvia urticifolia L. Rosa humilis Marsh.

Rhus t~hina ·L. Sanguinaria canadensis L. Ribes Groasularia L.* Ribes oxycanthoides L.* SanicUla canadensis L.

Rob·inia Pseudo-Acaoia L. SatureJa Nepeta (L.) Scheele. Satureja vulgaris (L.) Fritsch. Rubus ·odoratus L. Rubus villosus Ait. Saxifrage. virginiensis Miohx.

Rudbeckia hirta L. Sorophularia marilandica L. Rudbeckia laciniata L. Scutellaria lateriflora L. Ruellia cilosa Pursh. Scutellaria parvula Miohx. Scuteilaria pilosa Michx. - Rumex Acetosella L. Sedum ternatum Michx. Seneoio Balsamitae Muhl. Sonohus asper (L.) Hill. Senecio obovatus Kuhl. Sonchus oleraaeus L. var.elongatua {Pu.rah.) Britton.*

Sida Spinosa L. Sorghastrum nutans (L.)Nash.

Siline antirrhina L. Sp·ecularia perfoliata (L.) A.DC. Siline pennsylvanica Michx.

Siline stellat~ (L.) Ait.f. ~ stacrhys cordata Riddell. Siline virginioa L. Staphylea trifolia L. Silphium trifoliatum L.

Steironema ciliatum (L.) Raf. Sisymbrium officinale (L.) Soop. Steironema lanceolatum (Walt.) Gray. var.leiooarpum DC.

Si~yrinahium angustif olium Mill. Stellaria media (L.) Cyrill. Stellaria pubera Michx. Smilaoina racemosa (L.) Deaf. Stylosanthes biflora (L.) B.S.P. Smilax herbacea L.* Symphytum officinale L. Solanum aarolinense L.

S olanum nigrum L. Syringa vulgaris L. *

Solidago bioolor L. Solidago oaesia L.

I! Solidago Curtisii T.& G. Solidago flexioaulia L. Taraxaoum officinale Weber. Solidago hispida Muhl.* Solidago nemoralis Ait. Tephrosia virginiana (L.) Pers.* Teuorium aanaaense L. Vacoinium vaaillans Kalm.*

Thalictrum dioicum ~ L. Verbascum Blattaria L. Verbascum Thapsus L. Thuja oocidentalis L.* Verbena hastata L. Ti:tia amerioana L. Verbena urticaef olia L. Tilia europia L. Verbesina oocidentalis (L.) alt. Trichostema dichotomum L. Vernonia noveboracensis Willd. Tridens flavus (L.) Hitohc. ~

Veronica Anagallis~aquatioa L. Trif olium agrarium L. Veronica, of:ficinalis L. Trif olium arvense L. Veronica peregrina L. Trif olium hybridum L. Veronica serpyllif olia L. Trif olium incarnatum L. Veronica virginica L. Trif olium pratenae L. Trif olium procumbens L. Viburnum aoerif olium L. Trif olium repens L. . Viburnum prunif olium L. Viburnum rufidulum Raf. Trillium grandiflorum (Michx • .) - Salisb. Viola oanadensis L. Typha latifolia L.* Viola cucullata Ait. Viola fimbriatula Sm. Viola hastata Michxo Uvularia grandiflora Sm. Viola pedata L. Viola Rafinesquii Greene.* Viola rostrata Pursh. Viola soabriusou1a Schwein. Viola striata Ait.

Xanthium oanadense Millo

Zizia Bebbii {0oult.& Rose) Britton . :BIBLIOGRAPHY.

l ,. Arthur, Barnes, and Caul tor, • "Plnnt Dissection" . 2. Campbell, D. H. - "A University Text Book of Botany." 3. Chamberlain, C. J. - "llothods in Plant Histology." 4. Britton and Brol"lil - "!lluotrated Flora of the Northern States and Canada."

5. Goodale, G. L. - "Gray's Botanical Toxt Boo~" 1101. II. Physiological Botany • • 6. Gray, Asa, - "Field, Forest and Garden 3otany". ------"Gray's Botanical Text Book, Vol. I., Structural Botany." 8. Punnett, R. ·C. - "?.1endelisrn." 9. Robinson and Fernald - "Gray's New 11anual of Botany." 10. Stransburger, Noll, Sohonck, and Kersten - nA Toxt Book of Botany." 11. Thompson, J . A.- "The Science of Life". 12. Wilson, E. B. -"The Cell in Development and I nheritance."

13. Wood, A. - "Claas Bool~ of Botany. n ·