Using endophytes

to enhance golf's turfgrasses

Though much remains unknown, research programs are
making great strides in understanding how endophytes
can strengthen the resistance of turfgrass plants.
Mark Sellmann

E
ndoPhytes are beneficial fungi work to separate and identify the dif-
that live between the cell walls ferent races, strains and species of
of grass plants. Unlike fungi endophytes. Before endophytes can
such as bread molds or those causing be bred there has to be a way to tell
athlete's foot, fungal endophytes them apart - to tell the beneficial
don't cause an "irritation" reaction in ones from the unfavorable ones.
the plant. Rather, they actually help Possible benefits of using "cus-
the grass plant endure environmental tomized" endophytes are numerous.
stress conditions. For example, researchers are finding
Ironically, endophytes need stress that one of these benefits is drought
before they can shine. In other words, tolerance, which may prove invalu-
a turfgrass variety with endophytes able in regions where irrigation is re-
will not outperform one without stricted. University trials have also
endophytes under non-stressful con- found evidence of varying levels of
ditions. However, in stressful environ- enhanced resistance to chinch bugs,
ments such as drought, heat, sod webworms and white grubs
infection from Rhizoctonia, or feed- among varieties infected with differ-
ing of sod webworms, white grubs ent endophyte strains. Studies also
and chinch bugs, the variety with en- show better turfgrass recovery from
dophyte infection can tolerate and re- Endophyte fungi growing in sterile lab cul- diseases such as summer patch,
cover better than an endophyte-free ture. The one on the left is from the former
Yugoslavia, and the one on the right is
brown patch and dollar spot.
one. from Iran (bottom). Agrostis callus inocu- "A lot of present endophyte work
Today, more is known about the lated with endophytes in sterile lab culture for turfgrass purposes is being done
beneficial effects of endophytes in (top). Inoculation of callus tissue is one at universities such as Rutgers," Sun
turfgrass varieties than ever before. In method researchers use to put endo- says. "Rutgers had a very strong turf-
phytes into turfgrass.
addition, researchers are also zeroing grass program for a long time, so it's
in on some additional benefits they looked into enhanced insect resis- logical to have a well-established en-
envision for turfgrasses with endo- tance in fine fescues, ryegrasses, tall dophyte program there as well."
phytic fungus. fescues and Poa species. Researchers at other universities
Sun has worked to identify differ- are also conducting endophyte work.
Can made-to-order ent races of endophytes. "Most endo- James White at Auburn University at
endophytes be bred? phytes," he says, "have only an Montgomery has been screening the
Suichang Sun, former endophyte asexual (non-reproducing) stage. distribution of different endophyte
researcher at Rutgers University and They do not reproduce sexually. So species and has been collecting sev-
now on staff at Jacklin Seed Co., it's been a big debate over how those eral native endophytes.
screened turfgrass seeds and plant tis- endophytes should be handled. By White has been working in cooper-
sues for endophytes within turf vari- looking at the morphological charac- ation with the breeding program at
eties and germplasms. In the field he teristics of endophytes, we could only Rutgers under the direction of Dr.
get a rough idea of what real differ- Reed Funk. He has collected endo-
Mark Sellman is an associate plant
ences they have." phyte-infected grasses from Europe
breeder at Jacklin Seed Co. in Post
Researchers are currently perform- and is planning to travel to South
Falls, Idaho.
ing DNA tests and fingerprinting America in search of others.

62 Calf Course Management / June 7994

 Fungal endophytes grow within the cell walls between the host plant cells. Although most endophytes are fungi (above), bacterial endophytes
may also playa future role in the disease and insect resistance of their hosts.
He has been trying to determine
the range of the different species and
races of endophytic fungi. Might it be
possible that one race of endophytes
may give a plant more resistance to
an insect than another? White's work
suggests this may be true. His studies
also will determine that, because
there are multiple races of insects, it
may be possible that one particular
endophyte is more resistant to a par-
ticular race of insect than another.
Chuck West, an endophyte re-
searcher at the University of
Arkansas, is working with tall fescue
endophytes. He began his work by
looking at the favorable effects of fun-
gal endophytes on drought resistance
and has been trying to identify the
physiological mechanism involved.
"We're trying to identify strains of
endophytes that we can introduce
into some improved cultivar of tall
fescue that would enhance its ability
Golf Course Management / June 7994
to persist during drought, and insect
and nematode attacks," says West.
His work involves screening endo-
phytes from around the world for
their ability to handle different
stresses.
Bacterial endophytes
Generally, when "endophyte" is
mentioned, the reference is to fungal
endophytes as defined earlier. How-
ever, not all endophytes are fungal.
Some are bacterial, although these
endophytes have not been studied
nearly as much as their fungal
counterparts.
"The endophytes most people
have heard of are the fungal endo-
phytes," Sun says. "But there are also
endophytes that are bacterial. That's
a totally different story.
"Most fungal endophytes produce
alkaloids that give enhanced resis-
tance to insects and diseases. But
bacterial endophytes basically just live
in the plant."
At Rutgers, John Chen in the de-
partment of plant pathology is look-
ing closely at bacterial endophytes.
He found a bacterial endophyte in
bermudagrass while working in Tai-
wan. Chen is trying to introduce pes-
ticide-producing genes into the
bacteria rather than changing the
plant itself. The bacteria grow very
quickly within the plant and express
any genes transplanted by the scien-
tists.
Some work has been done with
bacterial endophytes in other crops
such as pearl millet and corn. Nick
Hill, endophyte specialist at the Uni-
versity of Georgia, found there are
even nitrogen-fixing bacterial endo-
phytes, although they are difficult to
work with. Hill claims that these en-
dophytes are more subject to the nu-
tritional whims of the plant.
Continued
63
ENDOPHYTES from p. 63 do have merit in the future, would and his colleagues stated in a recent
have several practical uses. One ad- journal article that although drought
In the future, bacterial endophytes vantage is that it might be possible to resistance has been reported for en-
will be used to carry and transport ge- spray bacterial endophytes onto a dophyte-infected tall fescue, no one
netic information into the plant. "Re- field or lawn. Fungal endophytes can has looked at the effects or interac-
searchers can use bacterial only be introduced through breeding tions of tall fescue genotypes (vari-
endophytes as carriers to transfer or inoculation of individual plants at eties or lines). White decided to look a
genes into the plant," Sun says. "So the seedling stage. little closer. The team's objectives
basically, it's a totally different con- Sun, like Hill, agrees that some were to determine the effects of en-
cept than the fungal endophyte." promising research with bacterial en- dophyte infection on water relations
Bacterial endophytes, if they ever dophytes has already been done in and the survival of water-stressed tall
Table 1. Percent incidence of endo- other crops. fescue, and then determine how they
phytes in seeds of cultivars of tall "What researchers are trying to do relate to the genetics of the plant.
fescues from the 1992 National Turf- is introduce an insect-resistant gene White and his colleagues concluded
grass Evaluation Program. into bacterial endophytes and then in- that although improved drought resis-
(From Rutgers Turfgrass Proceedings, 1993) tance has been reported for endo-
oculate these bacterial endophytes
onto the surface of seed," says Sun. phyte-infected plants, their research
Cultivar Mean
By this means, the bacteria and did not confirm these findings. They
SR8400 96 gene enter together and infect a summarized that endophyte-infected
SR8200 92 seedling. They go into the plant and plants may have superior drought re-
Bonsai 91 then eventually the resistant gene will sistance resulting from improved
SR8010 91 express itself as the plant matures. drought avoidance traits — such as
KY-31 w/endo 89 Whether the offspring from this plant deeper rooting and greater root den-
Micro DD 59 will be endophyte-infected remains to sity — or better pest resistance,
Pixie 47 be seen. Sun thinks that once the en- rather than substantial endophyte im-
Phoenix 47 dophyte enters the plant it is there to pacts on the physiology of the plant.
SR8210 47
stay — however, the seed harvested White noted that recovery from
Bonsai Plus 41
Arid 39 from that plant may or may not con- drought stress was affected by tall fes-
Duke 34 tain the endophyte. cue variety but not by endophyte in-
Alamo 33 fection. They measured total tillers,
Rebel Jr. 33 Fungal endophytes tiller survival, plant survival and recov-
Anthem 33 and drought tolerance ery weights and found similar levels
Astro 2000 31 Dr. James Murphy, turfgrass man- for endophyte-infected and endo-
Montauk 26 agement specialist at Rutgers, has phyte-free plants. Thus, they con-
Safari 26 been trying to document drought re- cluded that endophyte infection does
Bonanza 25 sistance in tall fescue as mediated by not necessarily endow tall fescue with
Lancer 24 endophytes. As he wrote in a recent superior drought resistance.
Monarch 22
17
journal article:
Olympic II
Kittyhawk 14 "We found improved drought toler- Endophytes in bentgrass
Finelawn Petite 12 ance when you have the endophyte. Although endophytes are prevalent
Lexus 11 But there seem to be specific endo- within the fescues and ryegrasses, at
Guardian 10 phyte and plant interactions — which the present time there are no com-
Rebel-3D 8 means there's a lot of opportunity to mercial varieties of creeping bent-
Avanti 1 improve drought resistance in tall fes- grass containing endophytes.
Twilight 1 cue through manipulation of endo- "So far we haven't found any endo-
Shenandoah 1 phytes and plant genotypes in phytes in creeping bentgrass and
Vegas 0 combination." colonial bent, but there are some
Austin 0 species of Agrostis that we know
Aztec 0 When subjected to drought stress,
Eldorado 0 endophytes and plants seem to work have endophytes in them," says Sun.
Silverado 0 synergistically to endure the stress. Other researchers are trying to in-
Tomahawk 0 Plants without endophytes under the oculate endophytes into creeping
Cochise 0 same conditions may not be able to bentgrass. Lisa Li, another researcher
Virtue 0 survive. at Rutgers, inoculated creeping bent-
KY-31 no endo 0 Endophytes in grasses improve re- grass with endophytes using tissue
Trailblazer II 0 sistance to insects, spur growth and culture. She did this by inoculating
Leprechaun 0 improve resistance to high tempera- cells of bentgrass with the fungus and
Falcon 0 ture and drought stress. At Texas then regenerating full-sized plants
A&M University, Dr. Richard White from the individual cells. The plants

64 Coif Course Management / June 1994

are now in the field. Chances are Program (NTEP) trial were infected high enough level to justify this desig-
good that bentgrass varieties will be with endophytes. nation. Sun and Smith went on to ex-
endophyte-enhanced in the future. Seventeen of the 26 chewings fes- amine the percentage of viable
The University of Georgia's Hill cues, nine of 14 hard fescues and endophytes in the entries. They saw
stresses the need to go back to the seven of 15 strong creeping red fes- this as very important because studies
centers of origin of the Agrostis genus cue entries were found to have endo- in the past have suggested that the vi-
to locate the original bentgrass endo- phytes in the seeds. No sheep or ability of endophytes in fine fescue
phytes and screen them for their pos- slender creeping red fescues were seed can dramatically decline in less
sible contributions. "Bentgrasses are found to be infected. than one year's storage at room tem-
European — they're everywhere More than half of the infected en- perature. Eventually customers may
from the Mediterranean all the way tries (17 of 33) were considered to be want to know not only whether the
up into England and France, and all highly infected (> 75-percent infec- variety is endophyte enhanced, but
through that area." tion). One figure worth noting is that also whether the endophytes are vi-
Is the best place to find endophytes only 22 percent of the entries in the able.
in Europe or are there also endo- 1989 NTEP test had endophytes, Sun and Smith examined three
phytes in South America and Africa? compared with 56 percent of the en- plants that grew from individual seeds
"From the studies I have done so far," tries in the 1993 NTEP test. That's a of each variety to determine the level
says Sun, "it seems that endophytes significant trend that will likely in- of viable endophytes. In many in-
in grasses are more common than we crease in the future. stances, endophytic mycelia (thread-
originally thought. So really, any- * * * like fungal bodies) in the plant were
where there are native species, I think As we see more varieties released not observed until the second or third
there is a good chance to find endo- as "endophyte enhanced," it will be a plant was examined.
phytes there." challenge for the seed industry to Sun and Smith thought this was
The centers of origin of turfgrass keep the endophyte infection at a Continued on p. 68
species would be the first place to
look. These centers include Europe, Table 2. Percent incidence of endophytes in seeds of cultivars of perennial
Asia, the Far East, central South ryegrasses from the 1990 National Turfgrass Evaluation Program.
America and Africa. Few studies have (From Rutgers Turfgrass Proceedings, 1991)
been done searching for endophytes
Cultivar Mean Stallion 58
in Africa and Asia. Caliente 54
Hill stresses that besides collecting Affinity 97 Premier 50
new germplasm with endophytes, dif- Yorktown III 97 Entrar 47
ferent races should be screened for Advent 97 Prestige 43
their strengths and weaknesses. APM 97 Derby Supreme 38
"Based upon what we know with tall Express 97 Lindsay 37
fescue, ryegrass and endophytes, we Dandy 96 Charger 34
should evaluate different endophyte Seville 96 Envy 30
strains for the benefits we recognize, Syn-P 96 Pebble Beach 29
and make sure they are there," he Barrage++ 94 Barrage 28
Duet 93 Rodeo II 27
says. "If they are, then we should start Linn 18
Manhattan II (E) 93
developing some new management Target Fiesta II 15
92
strategies for integrated pest manage- Assure 92 Cutless 14
ment control. We need to come up Pleasure 92 Cowboy II 12
with new endophyte management Repell II 92 Danilo 6
protocols for golf course superinten- Riviera 91 Ovation 5
dents," he says. Pennant 91 Loretta 4
Considering that most superinten- Gettysburg 91 Pennfine 1
dents put a considerable amount of Pinnacle 90 Gator 1
Legacy 90 Danaro 1
their time and money into greens Repell Allegro 1
89
maintenance, a bentgrass endophyte SR4200 89 Patriot II 0
is certainly an exciting area for turf- Commander 88 Troubadour 0
grass management. Regal 86 Toronto 0
Fine fescue endophytes Saturn 85 Taya 0
Sherwood 74 Surprise 0
At Rutgers, Sun and Dirk Smith Competitor 71 Nomad 0
performed a simple study to deter- Accolade 70 Meteor 0
mine which fine fescue varieties have Equal 68 Goalie 0
endophytes. They found that 3 3 of Calypso 66 Cartel 0
the 59 entries examined from the Citation II 59
1993 National Turfgrass Evaluation

Golf Course Management / June 1994 65

ENDOPHYTE from p. 65 plants from sandstone cliffwalls, they Of the 69 species of Poa in the
identified F. rubra subsp. pruinosa. United States, about one-third of
The infection level of this population them have actually been found with
possible because mycelia of Acremo- was 37 percent. endophytes. However, Poa praten-
nium typhinum, the endophyte sis, Kentucky bluegrass, is not one of
species in fine fescues, are usually White formulated the following the-
ory to explain why endophyte-in- them.
lower in concentration than those of Through extensive turfgrass endo-
Acremonium coenophialum in tall fected individuals predominate on
rock surfaces: Rock ecosystems are phyte research, much more is known
fescue or Acremonium lolii in per- today about endophytes than was
ennial ryegrass, and thus are harder difficult to colonize because of limited
moisture and nutrients. However, known 10 years ago. Scientists know
to find. They also hypothesized that several of the benefits, but cannot yet
the mycelia concentration was even once colonized they are seldom dis-
turbed. explain physiologically how it all hap-
lower in the seedling stage, particu- pens inside the plant. There is a lot
larly during winter. Sun and Smith It is expected that some of the en-
dophytes that originate in these more that we need to know and un-
found that percentage of seed infec- derstand.
tion was usually higher than the viable ecosystems infect hosts and may pro-
duce stroma. In low moisture condi- Some of the primary questions to
endophyte levels in the resulting be answered in the future are:
plants. tions, infected red fescue plants may
have a selective edge over endo- • Will there someday be Kentucky
In another study of fine fescue en- bluegrasses with endophytes in them?
dophytes, White and his associates phyte-free plants.
• Willour putting greens be planted
performed an investigation on the Endophytes of the future with endophyte-enhanced bentgrass
sexuality of red fescue endophytes. As of yet there are no Kentucky that permits reduced pesticide use?
They found that most red fescue en- bluegrass varieties with endophytes. o
dophytes in North America were sex-
ual - in other words, they are Table 3. Percent incidence of endophytes in seeds of cultivars of fine fes-
capable of being bred. cues from the 1993 National Turfgrass Evaluation Program.
(From Rutgers Turfgrass Proceedings. 1994)
Prior research found that once the
stroma (a primitive flower) is formed,
Cultivar Species % Infected Seed
sexual reproduction may occur on its
surface. Unfortunately, for turfgrass Dawson Slender creep. 0
seed production stroma development Scaldis Hard 0
reduces seed yield and limits the use- Ecostar Hard 21
fulness of endophytes in seed produc- Rondo Strong creep. 0
tion. However, the stroma are Pamela Hard 0
important in the life cycle of endo- Medina Chewings 0
phytes and in the economics of seed Brigade Hard 0
production. Molinda Chewings 0
White conducted studies in England Jasper (E) Strong creep. 100
where many red fescues occur na- Victory (E) Chewings 94
tively to determine the levels of endo- Chewings 27
Bridgeport
phyte infection and their Nordic Hard 0
stroma-forming capability. Popula- Seabreeze Slender creep. 0
tions from hedge banks, stone walls Shademaster II Strong creep. 14
and cliff walls in England were found Shadow (E) Chewings 54
to be high in endophyte infection. Discovery Hard 87
White found that different levels of Tiffany Chewings 100
endophyte infection may be ex- Flyer Strong creep. 0
plained by distinct subspecies of red Jamestown II Chewings 89
fescue occurring in different habitats. Jamestown Chewings 0
One example is that on sand Darwin Chewings 0
dunes, the F. rubra subspecies are- Aruba Strong creep. 0
naria predominates. Of the 37 plants 82
Brittany Chewings
of this species they found, all were Spartan Hard 0
free of endophytes. Creeping red fes- Banner II Chewings 78
cue from hedge banks and stone Reliant II Hard 98
walls, however, was found to be Aurora w/endo Hard 59
highly infected with endophytes. The Cascade Chewings 0
endophytes were present in levels of Common Creep. Strong creep. 0
50 to 100 percent among the popu-
lations. When they looked at the

68 Golf Course Management / June 7994